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Neurons have recently emerged as essential cellular constituents of the tumour microenvironment, and their activity has been shown to increase the growth of a diverse number of solid tumours1. Although the role of neurons in tumour progression has previously been demonstrated2, the importance of neuronal activity to tumour initiation is less clear—particularly in the setting of cancer predisposition syndromes. Fifteen per cent of individuals with the neurofibromatosis 1 (NF1) cancer predisposition syndrome (in which tumours arise in close association with nerves) develop low-grade neoplasms of the optic pathway (known as optic pathway gliomas (OPGs)) during early childhood3,4, raising the possibility that postnatal light-induced activity of the optic nerve drives tumour initiation. Here we use an authenticated mouse model of OPG driven by mutations in the neurofibromatosis 1 tumour suppressor gene (Nf1)5 to demonstrate that stimulation of optic nerve activity increases optic glioma growth, and that decreasing visual experience via light deprivation prevents tumour formation and maintenance. We show that the initiation of Nf1-driven OPGs (Nf1-OPGs) depends on visual experience during a developmental period in which Nf1-mutant mice are susceptible to tumorigenesis. Germline Nf1 mutation in retinal neurons results in aberrantly increased shedding of neuroligin 3 (NLGN3) within the optic nerve in response to retinal neuronal activity. Moreover, genetic Nlgn3 loss or pharmacological inhibition of NLGN3 shedding blocks the formation and progression of Nf1-OPGs. Collectively, our studies establish an obligate role for neuronal activity in the development of some types of brain tumours, elucidate a therapeutic strategy to reduce OPG incidence or mitigate tumour progression, and underscore the role of Nf1mutation-mediated dysregulation of neuronal signalling pathways in mouse models of the NF1 cancer predisposition syndrome. Mouse models of NF1-associated optic pathway glioma show that tumour initiation and growth are driven by aberrantly high levels of NLGN3 shedding in the optic nerve in response to retinal neuron activity.

Neurons have recently emerged as essential cellular constituents of the tumor microenvironment, where their activity increases the growth of a diverse number of solid tumors. While the role of neurons in tumor progression has been previously demonstrated, the importance of neuronal activity to tumor initiation is less clear, particularly in the setting of cancer predisposition syndromes. In the Neurofibromatosis type 1 (NF1) cancer predisposition syndrome, in which tumors arise in close association with nerves, 15% of individuals develop low-grade neoplasms of the optic pathway (optic gliomas) during early childhood, raising the intriguing possibility that postnatal light-induced optic nerve activity drives tumor initiation. Here, we employ an authenticated murine model of Nf1 optic glioma to demonstrate that stimulation of optic nerve activity increases optic glioma growth, while decreasing optic nerve activity via light deprivation prevents tumor formation and maintenance. By manipulating environmental light to modulate optic pathway (retinal) neuron activity, we show that Nf1 optic glioma initiation depends on neuronal activity during a developmental period susceptible to tumorigenesis. Germline Nf1 mutation in retinal neurons results in aberrantly high optic nerve neuroligin-3 (Nlgn3) shedding in response to retinal neuronal activity. Moreover, genetic Nlgn3 loss or pharmacologic inhibition of Nlgn3 shedding blocks murine Nf1 optic gliomagenesis and progression. Collectively, these studies establish an obligate role for neuronal activity in the development of certain brain tumors, elucidate a therapeutic strategy to reduce optic glioma incidence or mitigate tumor progression, and underscore the role of Nf1 mutation-mediated dysregulation of neuronal signaling pathways in the NF1 cancer predisposition syndrome.

Insulin-degrading enzyme (IDE, insulysin) is the best characterized catabolic enzyme implicated in proteolysis of insulin. Recently, a peptide inhibitor of IDE has been shown to affect levels of insulin, amylin, and glucagon in vivo. However, IDE(-/-) mice display variable phenotypes relating to fasting plasma insulin levels, glucose tolerance, and insulin sensitivity depending on the cohort and age of animals. Here, we interrogated the importance of IDE-mediated catabolism on insulin clearance in vivo. Using a structure-based design, we linked two newly identified ligands binding at unique IDE exosites together to construct a potent series of novel inhibitors. These compounds do not interact with the catalytic zinc of the protease. Because one of these inhibitors (NTE-1) was determined to have pharmacokinetic properties sufficient to sustain plasma levels >50 times its IDE IC50 value, studies in rodents were conducted. In oral glucose tolerance tests with diet-induced obese mice, NTE-1 treatment improved the glucose excursion. Yet in insulin tolerance tests and euglycemic clamp experiments, NTE-1 did not enhance insulin action or increase plasma insulin levels. Importantly, IDE inhibition with NTE-1 did result in elevated plasma amylin levels, suggesting the in vivo role of IDE action on amylin may be more significant than an effect on insulin. Furthermore, using the inhibitors described in this report, we demonstrate that in HEK cells IDE has little impact on insulin clearance. In total, evidence from our studies supports a minimal role for IDE in insulin metabolism in vivo and suggests IDE may be more important in helping regulate amylin clearance.

The species of the family Goodeidae have evolved reproductive strategies involving intraovarian gestation, early evacuation of nearly yolk-exhausted embryos from the ovigerous tissue into the ovarian cavity, placental matrotrophy during intraluminal gestation, and the birth of highly developed fry. The inner ovarian lining becomes hypervascularized during gestational periods and functions as the maternal component of the placental association. Embryotrophic liquid is secreted by the inner ovarian epithelium into the ovarian cavity. Comparative electrophoretic analyses of embryotrophe and maternal blood serum provide evidence for the transfer of maternal serum proteins into the embryotrophe. Trophotaeniae, proctodaeal processes of the embryos, provide a surface for nutrient absorption. Endocytic activity was demonstrated by ingestion of unspecific tracer proteins in various species. Moreover, the trophotaenial absorptive cells (TACs) in Ameca splendens ingest various proteins or random copolymers conjugated to colloidal gold as well as radioiodinated proteins in a way that satisfies the criteria of receptor-mediated endocytosis. Several aminopeptidases (APs) on the surface of TACs were identified as protein binding sites as evidenced by inhibition of binding and uptake of marker proteins in the presence of AP substrates or AP inhibitors. Morphological adaptations of the embryonic circulatory system pertaining to nutrient and gas exchange were characterized. The embryonic epidermis comprises two layers of squamous cells closely underlain by a dense capillary net. Efficient gas exchange is facilitated by a thin embryotrophe-blood barrier of both the embryonic skin and the intraovarian lining.

Paullinia cupana (guaraná) is an Amazonian plant that has been previously shown to be effective in treating chemotherapy-related fatigue (CRF) in patients with breast cancer. We aimed to evaluate the efficacy of a purified dry extract of P. cupana (PC-18) in patients with various solid tumors treated with chemotherapy.We included 40 patients with solid tumors who showed increases in their Brief Fatigue Inventory (BFI) questionnaire scores after 1 week of systemic chemotherapy. PC-18 was administered at 37.5 mg by mouth two times per day (PO bid), starting after 1 week of chemotherapy, for 3 weeks (induction phase). Patients who had an improvement in or stabilization of their BFI scores were randomized to receive either PC-18 at the same dose or placebo for the following 3 weeks (maintenance phase).After PC-18 treatment, the BFI fatigue scores improved or stabilized in 36 out of the 40 patients (mean BFI score difference = 2.503; 95% confidence interval: 1.716-3.375, p = .0002). Three weeks after randomization (16 patients on PC-18 and 17 on placebo), we observed no significant differences in the BFI, Functional Assessment of Chronic Illness Therapy, Hospital Anxiety and Depression Scale, and Pittsburgh Sleep Quality Index scores between patients randomized to PC-18 versus placebo.We conclude that the PC-18 extract may be effective for the treatment of CRF in patients with a variety of solid tumors. A conditioning effect, which was observed in patients who had a beneficial effect of PC-18 on CRF, may explain the better than expected fatigue scores of the placebo-treated patients.

A series of N-aryl pyridinone inhibitors of p38 mitogen activated protein (MAP) kinase were designed and prepared based on the screening hit SC-25028 (1) and structural comparisons to VX-745 (5). The focus of the investigation targeted the dependence of potency and metabolic stability on the benzyloxy connectivity, the role of the C-6 position and the substitution pattern on the N-phenyl ring. Further optimization produced the highly selective and potent pyridinones 2 and 3. These inhibitors exhibited activity in both acute and chronic models of inflammation.

Indoleamine 2,3-dioxygenase 1 (IDO1) is a heme-dependent enzyme that catalyzes the initial and rate-limiting step of tryptophan catabolism resulting in the local depletion of tryptophan and the concomitant production of kynurenine, both of which are immunosuppressive. Targeting IDO1 in combination with PD-1/PD-L1-targeted antibodies has shown promise in early phase clinical trials in several cancers and strongly suggests that, in some patients, IDO1 expression restrains PD-1/PD-L1-targeted checkpoint therapies. While some cancers extrinsically express IDO1 in response to IFN-γ produced from an ongoing, yet ineffective immune response, others select for the intrinsic expression of IDO1, independent of an immune response. We identified several cancer cell lines that intrinsically expressed either IDO1 or the related isozyme TDO2. Using these cell lines, we discovered LY3381916, a potent and selective inhibitor of cell-based IDO1 activity (IDO1 7 nM; TDO2 >20 µM). Using a variety of techniques, we demonstrated that LY3381916 binds to newly synthesized apo-IDO1 lacking heme, but does not inhibit mature heme-bound IDO1. Protein x-ray crystallography confirmed that LY3381916 binds to apo-IDO1 where it occupies the heme-binding pocket of IDO1. As a result of this novel mechanism of action, substantial inhibition of IDO1 in tumors requires the turn-over of mature heme-bound IDO1. Modeling of the pre-clinical PK/PD relationship suggests QD dosing of LY3381916 will maintain greater than 90% inhibition over 24 hours. In addition, due to the favorable properties of the drug, significant central nervous system (CNS) penetration has been measured for LY3381916 (rodent kp,uu 0.26). Kynurenine-mediated agonism of the aryl hydrocarbon receptor (AHR) is immunosuppressive in the tumor microenvironment. Inhibition of IDO1 and the subsequent reduction of kynurenine can relieve this immunosuppression. However, several heme-binding IDO1 inhibitors have been shown to replace kynurenine as an AHR agonist potentially limiting their ability to relieve this IDO1-dependent immunosuppressive mechanism. LY3381916 shows no confounding agonism of AHR up to 100 µM. Additionally, we characterized LY3381916 in pre-clinical tumor models and demonstrated that it was able to enhance LY3300054, anti-PD-L1 antibody (LY3300054) activity, which was associated with an enhanced T cell response. Based on these characteristics, LY3381916 is currently being investigated in a Phase I clinical trial. These data suggest further development of LY3381916 may be warranted. Citation Format: Frank C. Dorsey, Karim A. Benhadji, Lillian L. Sams, Debra A. Young, John F. Schindler, Karen L. Huss, Alexander Nikolayev, Carmine Carpenito, David Clawson, Bonita Jones, Andrew L. Faber, James E. Thomas, Steven A. Haney, Gaiying Zhao, William T. McMillen, Tod Smeal, Daniel J. Sall, Michael D. Kalos, Sandaruwan Geeganage, James R. Henry. Identification and characterization of the IDO1 inhibitor LY3381916 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5245.

Activation of the p38 kinase pathway in immune cells leads to the transcriptional and translational regulation of proinflammatory cytokines. Mitogen-activated protein kinase-activated protein kinase 2 (MK2), a direct downstream substrate of p38 kinase, regulates lipopolysaccharide (LPS)-stimulated tumor necrosis factor alpha (TNFalpha) and interleukin-6 (IL-6) production through modulating the stability and translation of these mRNAs. Developing small-molecule inhibitors of MK2 may yield anti-inflammatory efficacy with a different safety profile relative to p38 kinase inhibitors. This article describes the pharmacologic properties of a benzothiophene MK2 inhibitor, PF-3644022 [(10R)-10-methyl-3-(6-methylpyridin-3-yl)-9,10,11,12-tetrahydro-8H-[1,4]diazepino[5',6':4,5]thieno[3,2-f]quinolin-8-one]. PF-3644022 is a potent freely reversible ATP-competitive compound that inhibits MK2 activity (K(i) = 3 nM) with good selectivity when profiled against 200 human kinases. In the human U937 monocytic cell line or peripheral blood mononuclear cells, PF-3644022 potently inhibits TNFalpha production with similar activity (IC(50) = 160 nM). PF-3644022 blocks TNFalpha and IL-6 production in LPS-stimulated human whole blood with IC(50) values of 1.6 and 10.3 microM, respectively. Inhibition of TNFalpha in U937 cells and blood correlates closely with inhibition of phospho-heat shock protein 27, a target biomarker of MK2 activity. PF-3644022 displays good pharmacokinetic parameters in rats and is orally efficacious in both the rat acute LPS-induced TNFalpha model and the chronic streptococcal cell wall-induced arthritis model. Dose-dependent inhibition of TNFalpha production in the acute model and inhibition of paw swelling in the chronic model is observed with ED(50) values of 6.9 and 20 mg/kg, respectively. PF-3644022 efficacy in the chronic inflammation model is strongly correlated with maintaining a C(min) higher than the EC(50) measured in the rat LPS-induced TNFalpha model.

Nuclear factor (NF)-kappaB activation has been clearly linked to the pathogenesis of multiple inflammatory diseases including arthritis. The central role that IkappaB kinase-2 (IKK-2) plays in regulating NF-kappaB signaling in response to inflammatory stimuli has made this enzyme an attractive target for therapeutic intervention. Although diverse chemical classes of IKK-2 inhibitors have been identified, the binding kinetics of these inhibitors has limited the scope of their applications. In addition, safety assessments of IKK-2 inhibitors based on a comprehensive understanding of the pharmacokinetic/pharmacodynamic relationships have yet to be reported. Here, we describe a novel, potent, and highly selective IKK-2 inhibitor, PHA-408 [8-(5-chloro-2-(4-methylpiperazin-1-yl)isonicotinamido)-1-(4-fluorophenyl)-4,5-dihydro-1H-benzo[g]indazole-3-carboxamide]. PHA-408 is an ATP-competitive inhibitor, which binds IKK-2 tightly with a relatively slow off rate. In arthritis-relevant cells and animal models, PHA-408 suppresses inflammation-induced cellular events, including IkappaBalpha phosphorylation and degradation, p65 phosphorylation and DNA binding activity, the expression of inflammatory mediators, and joint pathology. PHA-408 was efficacious in a chronic model of arthritis with no adverse effects at maximally efficacious doses. Stemming from its ability to bind tightly to IKK-2, as a novelty, we demonstrated that PHA-408-mediated inhibition of IKK-2 activity correlated very well with its ability to modulate the fate of IKK-2 substrates and downstream transcriptional events. We ultimately directly linked IKK-2 activity ex vivo and in vivo to markers of inflammation with the inhibitor plasma concentrations. Thus, PHA-408 represents a powerful tool to further gain insight into the mechanisms by which IKK-2 regulates NF-kappaB signaling and validates IKK-2 as a therapeutic target.

Plus de 30 ans de recul de chirurgie fonctionnelle nous permettent d’affirmer aujourd’hui que pres de sept a huit patients tetraplegiques sur dix peuvent se voir proposer une chirurgie du ou des membres superieurs. La mise en route d’une collaboration multidisciplinaire dans ce domaine ne s’improvise pas. La reeducation postoperatoire doit s’appuyer sur des programmes codifies que des equipes aguerries ont eprouves. Le dialogue permanent etabli entre le chirurgien et l’equipe de medecine physique vise a repondre au souci de raccourcir les durees d’immobilisation tout en prevenant le risque de ruptures, d’adherences et d’autres complications locales.

Affective mixed states in bipolar disorders are a current matter of scientific debate and represent a complex clinical picture with coexisting manic and depressive symptoms. Treatment of mixed states is regarded as an important challenge. As diagnostic uncertainties complicate systematic clinical evaluations of this patient group, generally accepted clinical treatment guidelines are lacking yet. In this review the significance as well as the problems and risks of new treatment options are discussed.

The binding of N-cyclohexylformamide (CXF) to the complex of horse liver alcohol dehydrogenase with NADH mimics that of the Michaelis complex for aldehyde reduction catalyzed by the enzyme. The Raman spectra of bound CXF and its 13C- and 15N-substituted derivatives have been obtained using Raman difference techniques, and the results are compared with CXF spectra in aqueous solution and in methylene chloride. The results indicate that the amide N-H bond is trans to the C=O bond of CXF both in solution and in the enzyme ternary complex. The C=O stretch and N-H bending modes of the amide of CXF shift -16 and -9 cm-1, respectively, in the enzyme ternary complex relative to that in aqueous solution and -48 and 36 cm-1, respectively, relative to that in methylene chloride. Ab initio normal mode calculations on various model systems of CXF show that the observed frequency changes of the C=O stretch mode have contributions from the frequency changes induced by the environmental changes near both the local C=O bond and the remote N-H bond. The same is true for the observed N-H bending frequency change. Our calculations also show that the environmentally induced frequency changes are additive so that it is possible to determine the C=O stretch (or N-H bending) frequency change that is due to the local interaction change near the C=O (or N-H) bond from the observed frequency changes. On the basis of these results and the empirical relationship between the C=O stretch frequency shift and the interaction enthalpy change on the C=O bond developed here, it is found that the C=O group of CXF in the enzyme/NADH/CXF complex binds with a favorable interaction enthalpy of approximately 5.5 kcal/mol relative to water. Similar analysis suggests that the N-H moiety of CXF is destabilized in the ternary complex by about 1.5 kcal/mol relative to water but is stabilized by about 1.5 kcal/mol relative to a hydrophobic environment. The analysis describes quantitatively the binding of the C=O of CXF with the catalytic zinc and the hydroxyl group of Ser-48 and the interaction of the N-H with the benzene ring of Phe-93 of the enzyme.

The controlled study presented here was designed to examine the influence of the degree of structure of a coping-orientated therapy, the changes between the beginning and the end of therapy concerning satisfaction with therapy, psychopathology, attitude towards medication, coping behaviour and social variables, the effects on the days of hospitalisation within one year after the end of treatment and the relevant variables to predict the outcome twelve months later. It could be shown that patients in a relatively unstructured supportive group therapy wished to discuss topics that are very similar to those of coping-orientated therapy, that coping-orientated therapy results especially in an increase in knowledge about illness and treatment and that the therapeutic outcome after a one-year period can best be predicted by the degree of active coping strategies, social adaptation, trust in medication and cognitive variables.

Human alcohol dehydrogenase (HsADH) comprises class I (alpha, beta, and gamma), class II (pi), and class IV (sigma) enzymes. Selective inhibitors of the enzymes could be used to prevent the metabolism of alcohols that form toxic products. Formamides are unreactive analogues of aldehydes and bind to the enzyme-NADH complex [Ramaswamy, S.; Scholze, M.; Plapp, B. V. Biochemistry 1997, 36, 3522-3527]. They are uncompetitive inhibitors against varied concentrations of alcohol, and this makes them effective even with saturating concentrations of alcohols. Molecular modeling led to the design and synthesis of a series of cyclic, linear, and disubstituted formamides. Evaluation of 23 compounds provided structure-function information and selective inhibitors for the enzymes, which have overlapping but differing substrate specificities. Monosubstituted formamides are good inhibitors of class I and II enzymes, and disubstituted formamides are selective for the alpha enzyme. Selective inhibitors, with Ki values at pH 7 and 25 degrees C of 0.33-0.74 microM, include N-cyclopentyl-N-cyclobutylformamide for HsADH alpha, N-benzylformamide for HsADH beta1, N-1-methylheptylformamide for HsADH gamma2, and N-heptylformamide for HsADH sigma and HsADH beta1.

The substrate analogue L-aspartate beta-semialdehyde (L-ASA) has been identified as a mechanism-based inactivator of L-aspartase from Escherichia coli. The enzyme catalyzes the deamination of L-ASA to yield fumaric acid semialdehyde (FAA) and NH4+, with the product FAA partitioning between subsequent release or irreversible enzyme inactivation. Complete protection against L-ASA inactivation is observed in the presence of the product fumarate and a divalent metal ion. However, protection against inactivation by the product FAA also requires the presence of an enzyme activator. In addition to functioning as a mechanism-based inactivator, L-ASA has also been shown to serve as an activator of L-aspartase. The mechanism of inactivation by FAA involves the attack of an active site nucleophilic at the alpha-carbon of FAA to yield a stable Michael type enzyme adduct. Subsequent formation of a hydrazone upon treatment of the enzyme adduct with 2,4-dinitrophenylhydrazine confirms the presence of the unreacted aldehydic group of FAA. Examination of a group of product analogues with different substituents has demonstrated a correlation between the electron-withdrawing ability of these functional groups and the rate of inactivation of L-aspartase.

Teleosts do not possess uteri and ovulation occurs into the ovarian lumen. Hence, osteichthyan viviparity is characterized by intrafollicular gestation or intraluminal gestation (within the ovarian cavity), and thus, the ovary functions both in egg production and maintenance of the developing embryos. Because there is no attachment between embryonic and maternal tissues, maternal-embryonic exchange of gas, electrolytes, nutrients, metabolic waste, and possibly effector molecules and immunoglobulins is mediated by embryotrophe. In several instances it has been demonstrated that embryotrophic proteins originate from maternal serum rather than from elaboration within ovarian tissues. In intraluminal gestation, the ovarian wall becomes hypervascularized and the inner ovarian epithelium lining the ovarian cavity secretes embryotrophe. Trophotaeniae provide extraembryonic endodermal exchange surfaces in many species undergoing intraluminal gestation. There is a strong indication of a trophotaenial membrane binding system for proteins containing appreciable amounts of lysine. In intrafollicular gestation, yolk, pericardial and coelomic sacs play a major role in physiologic exchange. In most instances, intrafollicular gestation is associated with extraembryonic fine structural modifications consistent with respiratory and osmoregulatory functions. The postulated role in nutrient transfer requires experimental evidence. In matrotrophic viviparous teleosts, the gut may provide an effective absorptive surface. On the other hand, the skin, and perhaps the gills and buccopharyngeal tissue, serve as respiratory surfaces. As yet, no experimental evidence suggests exchange of metabolites at these sites. © 1993 Wiley-Liss, Inc.

Objective: To compare the results for homocysteine concentration using chemiluminescence and HPLC methods in samples from school-age children. In addition, to determine the reference values for patients of this age group and assess the real prognostic value of homocysteine in healthy children. Methods: A prospective observational study was undertaken to determine plasma levels of homocysteine using two different assays, HPLC and chemiluminescence, in 185 samples from school-age children living in Santo Andre, with no chronic or inflammatory diseases, and absence of pubertal development. Results: The results were presented in percentiles and reference values were determined within this age group (7-9 years old). Homocysteine concentration ranged from 2.0 to 9.9 µmol/l (r = 0.821 and p < 0.001). Conclusions: It was verified that chemiluminescence is comparable to HPLC when both techniques are used to detect homocysteine in school-age children. There is an important correlation between both methods, which allows investigation of this amino acid as a risk factor for heart diseases.

Nuclear factor-kappaB (NF-kappaB) is one of the major families of transcription factors activated during the inflammatory response in asthma and chronic obstructive pulmonary disease. Inhibitory factor-kappaB kinase 2 (IKK-2) has been shown to play a pivotal role in cytokine-induced NF-kappaB activation in airway epithelium and in disease-relevant cells. Nevertheless, the potential toxicity of specific IKK-2 inhibitors may be unacceptable for oral delivery in chronic obstructive pulmonary disease. Therefore, local delivery to the lungs is an attractive alternative that warrants further exploration. Here, we describe potent and selective small-molecule IKK-2 inhibitors [8-(5-chloro-2-(4-methylpiperazin-1-yl)isonicotinamido)-1-(4-fluorophenyl)-4,5-dihydro-1H-benzo[g]indazole-3-carboxamide (PHA-408) and 8-(2-(3,4-bis(hydroxymethyl)-3,4-dimethylpyrrolidin-1-yl)-5-chloroisonicotinamido)-1-(4-fluorophenyl)-4,5-dihydro-1H-benzo-[g]indazole-3-carboxamide (PF-184)] that are competitive for ATP have slow off-rates from IKK-2 and display broad in vitro anti-inflammatory activities resulting from NF-kappaB pathway inhibition. Notably, PF-184 has been designed to have high systemic clearance, which limits systemic exposure and maximizes the effects locally in the airways. We used an inhaled lipopolysaccharide-induced rat model of neutrophilia to address whether inhibiting NF-kappaB activation locally within the airways would show anti-inflammatory effects in the absence of systemic exposure. PHA-408, a low-clearance compound previously shown to be efficacious orally in a rodent model of arthritis, dose-dependently attenuated inhaled lipopolysaccharide-induced cell infiltration and cytokine production. Interestingly, PF-184 produced comparable dose-dependent anti-inflammatory activity by intratracheal administration and was as efficacious as intratracheally administered fluticasone propionate (fluticasone). Together, these results support the potential therapeutic utility of IKK-2 inhibition in inflammatory pulmonary diseases and demonstrate anti-inflammatory efficacy of an inhaled IKK-2 inhibitor in a rat airway model of neutrophilia.

In an attempt to determine the role of vitamin E in retrolental fibroplasia (RLF) we report our experience with 191 infants of less than 1500 g birth weight. Of these infants, 16.75% had evidence of acute RLF in hospital, 8.4% had cicatricial RLF at follow-up, and four infants (2.1%) were blind, none of whom had received supplementary vitamin E. The incidence of cicatricial RLF at follow-up was significantly lower in infants who received vitamin E early after birth (12 h) than in those who did not (3 of 105 versus 13 of 86, X2 = 9.26, P = 0.002), as was the incidence of Grade III or greater cicatrix (0 of 105 versus 7 of 86, X2 with Yates = 6.72, P = 0.01). Stepwise multiple linear regression analysis revealed three factors distinguishing infants who developed cicatricial RLF from those who did not: the lack of early vitamin E supplements (P = 0.0023), the significantly larger number of arterial PO2 values over 100 mmHg (P = 0.0056), and the presence of an intraventricular haemorrhage (P = 0.0032). The incidence and severity of necrotizing enterocolitis was similar in infants who received vitamin E and in those who did not. It is recommended that vitamin E be given within the first 12 hours of birth to all infants of less than 1500 g who require supplementary oxygen.

Alkaline phosphatase (AP) has several isoforms including bone alkaline phosphatase (BAP). We evaluated BAP and AP for screening for bone metastasis (BM) in patients with solid tumours. This is a prospective non-blinded study conducted at ABC Foundation School of Medicine Oncology clinics. A total of 40 subjects without a history of cancer and 62 patients with various solid tumours referred for a bone scan had serum drawn for BAP and AP determination. Bone alkaline phosphatase and AP levels in patients with cancer and BM, without BM and with no cancer, were 70.32 +/- 3.65 and 310.21 +/- 16.87 U/L; 41.40 +/- 2.80 and 113.23 +/- 12.95 U/L; 21.19 +/- 2.76 and 148.05 +/- 12.79 U/L respectively (P < 0.0001 for both AP and BAP). For BAP and AP sensitivity, specificity, positive and negative predictive values were 0.86 and 0.52; 0.69 and 1; 0.45 and 1; 0.94 and 0.87 respectively. ROC AUC value for BAP was 0.89 and for AP was 0.93. We conclude that BAP is more sensitive than AP, whereas AP had a remarkable specificity of 100%. In screening for BM in patients with solid tumours, obtaining initially BAP and then selecting for further investigation only patients with an abnormal AP may be a cost and resource saving strategy.

International audience; Study design:Retrospective study.Objectives:To determine the potential impact of rehabilitation care on associated symptoms and functional improvements of paraplegic patients with metastatic spinal cord compression.Setting:CMN Propara, Montpellier (France).Measures:Demographics, Functional Independence Measure (FIM), Frankel Modified Score and Visual Analog Scale (VAS) for pain, intercurrent adverse medical events and neurological outcome, duration of stay, survival time, rehospitalization in a non-Spinal Cord Injury unit, number of contracts defining the patients rehabilitation goals, number of contracts defining the patients duration of stay within the rehabilitation center.Results:We reviewed the charts of 26 patients. The initial neurological profile was paraplegia or paraparesis for 24 patients and quadriparesis for 2 patients. Regarding functional improvements: four patients demonstrated a poor functional evolution, five patients showed no functional improvements or very slight improvements and all the other patients showed an increase in their overall functional aptitudes. At the end of the stay, 14 patients were urinary independent. Our study reports 52 rehospitalizations in an another unit and 101 outpatient visits during their rehabilitation stay in a physical medicine and rehabilitation (PM&R) center. For the 14 patients who were deceased at the time of data collection, the median survival rate post-paraplegia was 12.7 months. A total of 12 of the 14 patients spent more than a third of their remaining survival time in a rehabilitation center.Discussion:Compared to the patients' life expectancy, their stay in a rehabilitation center is too long and prevents them from spending time with family and loved ones. The occurrence rate of the associated symptoms is high because of both cancer-related disorders and neurological disorders caused by the spinal cord lesion. PM&R professionals are faced with patients affected by chronic pain and fatigue as well as frequent rehospitalizations, short stays and outpatient stays, in the primary oncology unit. This study focuses on the need to privilege the patients' comfort over their functional rehabilitation.Spinal Cord advance online publication, 10 June 2008; doi:10.1038/sc.2008.75.

Gravid goodeid females harbor embryos in a preformed ovarian cavity for prolonged periods of gestation. Various nutrients for embryonic growth are provided by the internal ovarian epithelium (IOE). Its cells flatten during late stages of gestation and form an attenuated layer of cytoplasm covering a dense network of protruding capillaries, with the nuclear domains mostly recessing between the vascular meshes. The IOE in both Xenotoca eiseni and Girardinichthys viviparus exhibit morphological features associated with vesicular transport of macromolecules. The amounts of rough endoplasmic reticulum in the IOE cells seem insufficient to effectively synthesise proteinaceous secretions. Apparently, it rather serves as a transit route for serum-derived products. Cationized ferritin (CF) was injected into the ovarian cavity of gravid females. The electrostatic ligand spotwise attached to the luminal surface of the IOE and gained access by adsorptive micropinocytosis. Many tracer molecules were sequestered into lysosome-like vacuoles that became increasingly swollen after prolonged incubation intervals. In addition, CF traversed the IOE within small vesicles. At the basal pole of the cells the contents of transcytotic vesicles were evacuated, and localization of small CF-clusters was regularly in the basement lamina, in the underlying connective tissue, in vacuoles within migrant cells, in vesicular compartments of the capillary endothelia, in capillary lumina, and in intravascular leucocytes. Tracer molecules were never observed to enter stacked Golgi cisternae. Since the cationic marker probably follows retrograde pathways of the protein secretion, the experimental data support the morphologically derived conclusions that postulate a major role for the IOE in transepithelial transport.

The trophotaenial absorptive cells (TACs) in goodeid embryos facilitate nutrient absorption during prolonged periods of intraovarian gestation. In a study of membrane differentiations associated with solute and ligand transfer in the trophotaeniae of Xenotoca eiseni, embryos were incubated in vivo with cationized ferritin (CF) prior to freeze-cleaving. This exposure to high concentrations of an adsorptive ligand was meant to induce swelling of the endosomal compartment. Macromolecular trafficking in TACs occurs via an apical endocytic complex consisting of plasma membrane invaginations, a large population of small vesicles, uniformly thick apical tubules, and endosomes. Freeze-fracture replicas showed that the microvillar plasma membrane P-face of TACs was studded with intramembrane particles (IMPs) at a fairly high density, whereas that of the cell surface proper contained a distinctly lower density and the tubulovesicular endocytic pits contained almost no IMPs. The majority of small vesicles and apical tubules in a near surface position displayed P-fracture faces with only a few odd IMPs, indicating that membrane, shuttling between the apical plasma membrane and intracellular sorting organelles, obviously does not carry along many large-sized integral membrane proteins. The distended endosomal compartment had many P-face-associated particles primarily clustered into patches. Specializations of the lateral plasma membrane included 4–8 tight junctional strands, relatively large complements of gap junction proteins, and numerous plaques of desmosomal membrane particles. A system of lamellar cisternae underlay the lateral cell surface that was in continuity with the intraepithelial space by numerous tubular canals, giving rise to an intracellular amplification of the basolateral plasma membrane. Their outward openings appeared as tiny pits on the cytoplasmic faces of freeze-cleaved cell membrane. The density of IMPs on the P-faces of the surface plasma membrane was apparently lower than that on its invaginated lamellar complex. Hence, it is concluded that the mobility of integral membrane proteins in the plane of the membrane may be hampered in movement across the surface pores.

Using PET with the opioidergic ligand [11C]diprenorphine, the authors demonstrate decreased tracer binding in the pineal gland of cluster headache patients vs healthy volunteers. Opioid receptor availability in the hypothalamus and cingulate cortex depended on the duration of the headache disorder. Therefore, the pathophysiology of cluster headache may relate to opioidergic dysfunction in circuitries generating the biologic clock.

The kinetic mechanism of mitogen-activated protein kinase activated protein kinase-2 (MAPKAPK2) was investigated using a peptide (LKRSLSEM) based on the phosphorylation site found in serum response factor (SRF). Initial velocity studies yielded a family of double-reciprocal lines that appear parallel and indicative of a ping-pong mechanism. The use of dead-end inhibition studies did not provide a definitive assignment of a reaction mechanism. However, product inhibition studies suggested that MAPKAPK2 follows an ordered bi-bi kinetic mechanism, where ATP must bind to the enzyme prior to the SRF-peptide and the phosphorylated product is released first, followed by ADP. In agreement with these latter results, surface plasmon resonance measurements demonstrate that the binding of the inhibitor peptide to MAPKAPK2 requires the presence of ATP. Furthermore, competitive inhibitors of ATP, adenosine 5'-(beta,gamma-imino)triphosphate (AMPPNP) and a staurosporine analog (K252a), can inhibit this ATP-dependent binding providing further evidence that the peptide substrate binds preferably to the E:ATP complex.

The isolated vascular effects of intravenous administration of the angiotensin converting enzyme (ACE) inhibitor enalaprilat were investigated. Thirty male patients undergoing cardiopulmonary bypass (CPB) were studied. According to a randomized sequence, 0.04 mg kg-1 enalaprilat (low-dose, n = 10), 0.08 mg kg-1 (high-dose, n = 10) enalaprilat or saline solution as placebo (control group, n = 10) was given as an i.v. bolus during CPB. Changes in mean arterial pressure (MAP) and venous reservoir (RV) of the extracorporeal circulation were studied as indices of arterial resistance and venous capacitance. Mean arterial blood pressure (MAP) and peripheral vascular resistance (SVR) were significantly more reduced in the high-dose enalaprilat group (MAP: -36 mm Hg after 9 min; SVR: -836 dyn s cm-5) than in the low-dose group (MAP: -13 mm Hg after 10 min). Volume of the reservoir (RV) decreased in both enalaprilat treated groups indicating additional (dose-dependent) venous pooling effects of the substance (low-dose: -300 ml; high-dose: -520 ml; control group: -100 ml). Skin capillary blood flow measured by laser Doppler flowmetry (LDF) increased after injection of 0.04 mg kg-1 enalaprilat, whereas it decreased significantly when MAP fell markedly in patients treated with high-dose enalaprilat. I.v. enalaprilat had dose-dependent vasodilating properties in the arterial and venous vessel system indicating reduction in pre- and afterload. Microcirculation in both enalaprilat treated groups improved as long as reduction in blood pressure was not limited.

There are reports of globulin abnormalities in various types of tumors during chemotherapy. Recently, even IgE levels seem to play an important role in the tumor's immune response. Trace elements, like Selenium (Se), act as basic components of essential system which play roles in immunology. Chemotherapy has been associated with decrease concentration of Se and this fact has been correlated with immunodepression states.

9102 Background: Hot flashes and sexual dysfunction often affect breast cancer's survivors compromising their quality of life. Approximately 65% of women after treatment for breast cancer complain ...

9017 Background: Chemotherapy-induced Nausea and Vomiting (CINV) is a distressing side-effect that affects many patients undergoing emetogenic chemotherapy, despite the use of antiemetic medication...

Single crystals of L-aspartate ammonia-lyase (L-aspartase) from Escherichia coli have been obtained by microdialysis at room temperature using polyethylene glycol 3350 and sodium acetate as co-precipitants. The crystals exhibit the symmetry of space group P2(1)2(1)2 with a = 156.5 A, b = 147.6 A, c = 102.5 A and diffract at least to 2.8 A.

Le trophotaenia et l'epiderme abdominal des embryons de Xenoophorus captivus ont ete etudies par microscopies optique et electronique et par des techniques histochimiques. Les specialisations morphologiques associees a la respiration cutanee ont ainsi ete examinees

Jenynsia lineata retains its embryos within the ovarian cavity for a prolonged gestation. In the absence of egg envelopes, maternal-embryonic transfer occurs through ovarian fluid across apposed epithelia, relatively lining the ovarian lumen and the surface of the embryos. There are no hypertrophied extraembryonic structures that could provide expanded exchange surfaces for the passage of nutrients beyond the 8-mm stage, but structural specializations of the ovary then form, and these may sustain embryogenesis. Outgrowths of the inner lining of the ovary, villi ovariales, enter the pharyngeal cavity of the embryos via an opercular cleft remaining from early stages of development, after depletion of yolk reserves, until shortly before term. The ovary and its villi are lined by a monolayer of squamous cells showing evidence of vesicular transport of macromolecular substances both on the apical surface and at the basolateral pole. It serves for transcellular passage of maternally derived substances rather than as a source of secretory products. Most adjacent cells interdigitate, and the epithelium is continuous except for few gaps at the villous tips, which allow paracellular passage of particulate matter. These epithelial cells contain abundant filaments, electron-dense granules within the cytoplasm and the nucleus, sparse elements of the rough endoplasmic reticulum, a Golgi apparatus, and different sorts of vacuoles. The capillaries in the intraovarian lining are spaced most densely at the ovarian wall, less so toward the tips of the villi. The villi ovariales contain a network of connective tissue that forms endotheliumlike septa, which divide the interior into numerous different-sized loculi.

Des echanges nutritionnels entre la mere et l'embryon de Xenoophorus captivus et les macromolecules proteiques du liquide nutritionnel ovarien ont ete etudies par focalisation isoelectrique du serum sanguin de l'embryon et de la mere. Le tissu ovarien a ete observe par microscopie electronique

Investigation of GFAP (Glial Fibrillary Acidic Protein) in 175 brain tumours showed varying amounts of fibrillary acidic protein in every glioma. In ependymal and oligodendroglial tumours a high number of positive neoplastic elements were detected, GFAP positive were also the peri-vascular cells of a so-called astroblastoma. In pilocytic astrocytomas, Rosenthal fibers were in part GFAP positive, in part negative. In giant cells gliomas, giant cells were GFAP negative or weakly positive. Intraleptomeningeal growing tumour cells presented usually a very strong positivity. In 8 recurring oligodendrogliomas, the number of GFAP positive tumour cells was the same in the primary tumour and in its recurrence. These results demonstrate that GFAP is not a specific astrocytic, but a glial-specific protein. Although GFAP is usually present in greater concentration in differentiated, slow growing gliomas, absolute reliable predictions on biological behaviour of the individual tumour are not possible, because a high GFAP content can be detected also in malignant tumours. GFAP investigation does not seem reliable for solving the pathogenetic problems of undifferentiated tumours: the results obtained in 50 medulloblastomas showed that the investigation of small tumour samples or the positivity of a single cell are inadequate data for a correct evaluation of the findings, especially bearing in mind that GFAP of degenerated astrocytes can be phagocytised by other cells, these findings giving rise to misinterpretations.

Carp (Cyprinus carpio) hemoglobin readily autoxidizes in blood smears. Quantification of Soret-band absorbance in individual erythrocytes by means of scanning cytophotometry therefore requires more elaborate methods of preparation of blood samples. Of the fixatives that have been tested, suspension of whole blood in isotonic salt solutions containing glutaraldehyde was most suitable. Glutaraldehyde-fixed red blood cells are totally resistant to hemolysis. In the course of fixation, hemoglobin is transformed to methemoglobin. Spectrophotometry indicated extensive similarities between glutaraldehyde-fixed carp methemoglobin and human methemoglobin. In aqueous solutions, the intensity of the Soret-peak was pH-dependent. The allosteric modifier organic polyphosphate caused an R----T transition, resulting in increased molar extinctions. Dried preparations showed Soret-spectra that were not influenced from either pH or organic polyphosphate concentration of the aqueous suspensions in which the erythrocytes had been stored. The same was true for slide preparations of cyanomethemoglobin, easily derived from methemoglobin on addition of potassium cyanide. In the absence of oxygen fresh blood cells from carp slowly transform their hemoglobin into deoxyhemoglobin. Spectra of the intermediate stages of deoxygenation, Hb4(O2)3, Hb4(O2)2 and Hb4(O2), as well as mixtures of these intermediates, could be monitored.

In an attempt to determine the role of vitamin E in retrolental fibroplasias (RLF), experience with 174 infants of less than 1500 g, all of whom survived beyond four weeks of age, the time of the initial eye examination, is reported. In Phase I, a prospective randomized controlled trial of parenteral vitamin E, 48 infants received intramuscular vitamin E and 6.3% had evidence of cicatricial RLF at follow-up, compared to 51 controls of whom 10% had cicatricial RLF. In Phase II, a nonrandomized prospective experience with the administration of oral vitamin E, 44 infants received early ( 2 = 5.75, P = 0.017), as was the incidence of Grade III or greater cicatrix (0 of 92 vs 7 of 82, χ 2 with Yates=6.12, P = 0.013). Stepwise multiple linear regression analysis revealed that two factors distinguished infants who developed cicatricial RLF from those who did not; the lack of early vitamin E ( P = 0.0000), and the number of days of exposure to supplemental oxygen ( P = 0.002). It is recommended that vitamin E be given within 12 hours of birth to all infants less than 1250 g who require supplemental oxygen.

Carp erythrocytes were fractionated by angle-head centrifugation which yielded fractions with a linear increase in density. Haematological examinations revealed that the heavier red blood cells of carp had greater volumes (MCV), more haemoglobin (MCH) and higher haemoglobin concentrations (MCHC) than light ones. The same experiments with human red cell fractions yielded a decrease in MCV, constant MCH and an increase in MCHC. Haemoglobin content in individual erythrocytes was also determined by scanning stage absorbance cytophotometry to establish the frequency distribution of the cellular haemoglobin contents. In carp, the distribution was symmetrical with the means increasing with density. No such change with cell density was found in human erythrocytes. Both carp and human erythrocytes incorporated [2-14C]glycine in vitro. After gel filtration, radioactivity was detected in carp, but not in human, haemoglobin fractions. 14C was found in all three haemoglobin fractions, obtained by isoelectric focusing, and was present in the haem and in the globin. [2-14C]glycine-labelled erythrocytes were reinjected into chronically cannulated carp and followed in vivo for several months. With time, the main peak of scintillation counts shifted from red cell fractions of low to high density. This is considered as evidence that density and age of red cells in carp are positively correlated and that erythrocytes can synthesize haemoglobin while circulating in the peripheral blood.

Scanning and transmission electron microscopy were used to examine the morphology of the perianal processes (trophotaeniae) of goodeid embryos (Girardinichthys viviparus) at two stages of gestation. The epithelial surface of trophotaeniae is composed of two cell types, one of which shows distinct features associated with absorptive activity. Such cells are characterized by microvilli, abundant mitochondria, and an agranular tubulolamellar network. Micropinocytosis at the apical surface is relatively rare. The brush border membranes contain high levels of alkaline phosphatase. The cells of the second type are the minor component of the trophotaenial epithelium. Their surface is distinct, due to the presence of microridges rather than microvilli. The reticulate arrangement of the cells gives rise to intercellular spaces which occasionally are very large. These interstices are populated with leukocytes. The histological appearance of these sections indicates that this tissue is involved in gas exchange. Embryos at very early stages of development possess similar epithelia which are differentiated to a lesser extent. The connective tissue in some parts of the processes shows structural modifications. It is densely packed with numerous leukocytes occupying the spaces between the cytyoplasmic ramifications of the stroma cells. Possible roles of the trophotaeniae in absorption, respiration, excretion, and the acquisition of immunity are discussed, and it is concluded that the perianal processes of the Goodeidae are more than just trophic embryonic structures.

Scanning stage absorbance cytophotometry was used to examine haemoglobin contents in individual cells or carp erythrocyte populations. Changes in frequency distributional profiles in response to respiratory stress caused experimentally by hypoxia and/or bleeding were studied at intervals. Histograms were drawn of haemoglobin contents of individual red blood cells in populations obtained from the same carp on four consecutive days after the fish was temporarily exposed to hypoxic environment. The modes and means of the haemoglobin values increased during the 3 days following to the stimulus, whereas a decline was measured on the 4th day. Erythrocyte counts showed the total number of red blood cells to be approximately constant during the period of investigation. On the basis of these findings, it is concluded that the mature, nucleated red blood cells of carp are capable of resuming haemoglobin synthesis after stimuli such as reducing ambient oxygen concentration. Frequency distributional profiles covering a period of 4 weeks following severe loss of blood showed that it took 10–12 days after bleeding until masses of immature erythrocytes appeared in the peripheral blood and that there were then two distinct populations of red blood cells. In the course of about 2 weeks the precursor cells developed into mature erythrocytes.

Zonae pellucidae of the viviparous goodeid teleosts Girardinichthys viviparus, Xenoophorus captivus, and Xenotoca eiseni were investigated ultrastructurally, and binding sites for ConA and WGA were localized on cross-sections using a colloidal gold technique. In late stages of development, the oocytes are surrounded by a three-zonated acellular matrix multiply perforated by pore canals allowing long microvilli of the oocyte to penetrate interstices of the follicle epithelium. Together, the surface of the microvilli and zona pellucida is coated by a thin layer of homogeneous slightly electron-dense material. In early oogenesis, the thin acellular layer is entirely packed with binding sites for WGA, whereas those for ConA occur only sparsely. Three-zonated zonae pellucidae amply contain both WGA and ConA receptors. The asymmetric labelling pattern obtained with both lectin protein gold preparations indicates a polarized organization of the different glycoconjugates. WGA receptors are concentrated within the outer region of the zona pellucida. Labelling with ConA-HRP-Au complexes produced heavy deposits of marker beads within the inner two thirds of the zona pellucida and weak labelling of the superficial coat. After prolonged digestion with neuraminidase, WGA binding sites were no longer detectable.

The endodermal trophotaenial epithelium in goodeid embryos acts as a placental exchange site. Fine structural and cytochemical data indicate that the trophotaenial absorptive cells are endocytotically highly active. To test their micropinocytotic capacity and characterize the cellular mechanisms involved in membrane, solute and ligand movements, living embryos of Xenoophorus captivus were incubated in saline media containing horseradish peroxidase (HRP) and/or cationized ferritin (CF) in vitro, and the uptake of these tracer proteins examined by both time sequence analysis and pulse-chase procedures. In some embryos, the effects of prolonged exposure to CF injected into the ovarian cavity, was also investigated. Labelling of the free cell surface was detectable with CF only, but interiorization of both probes was quick from all incubation media. Adsorptive pinocytosis of CF and fluid-phase uptake of HRP sequentially labelled pinocytic vesicles, endosomes, and lysosome-like bodies. In addition, CF-molecules were sequestered within apical tubules and small vesicles. HRP was largely excluded from both organelles and ended up in the lysosomal compartment. For CF, two alternative pathways were indicated by the pulse-chase experiments; transcellular passage and regurgitation of tracer molecules to the apical cell surface. The latter procedure involves membrane and receptor recycling, in which apical tubules are thought to mediate. In double-tracer experiments, using an 8:1 excess of HRP, external labelling with CF was light or lacking after 1-3 min, and the initial uptake-phase produced pinocytic vesicles and endosomes that mainly contained HRP-reaction product. Prolonged incubation, however, resulted in densely CF-labelled plasmalemmal invaginations and pinocytic vesicles that predominantly carried ferritin granules. After 60 min, the vacuoles of the endosomal compartment contained either high concentrations of HRP-reaction product, both tracers side by side, or virtually exclusively CF.

The effect of Vitamin E on the incidence and severity of RLF and BPD remains unclear. A prospective randomized controlled trial to further assess the role of Vit E in infants of 750-1500 gm to date includes 79 surviving infants (total n=105) of 29.5 wk (26-33 wk) gest., and birth weight 1130 gm (760-1500 gm). Randomization into groups was performed according to birth weight, (250 gm increments) initial Fi02 and need for intubation. Parenteral Vit E was administered on a fixed schedule within 12 hr of birth to 30 of the infants for 4 wk for a total dosage of 430 units followed by oral administration if tolerating feeds, or continued parenteral administration until 6 wk of age. There was no clinically significant difference between the two groups. Vit E levels were not significantly different at birth but were significantly higher in the Vit E group through-out the first 6 wk (33.8 vs. 6.9 mg/l, p