Strains of the Gram-negative bacterium Cronobacter (Enterobacter) sakazakii have been identified as emerging opportunistic pathogens that can cause enterocolitis, bacteraemia, meningitis, and brain abscess, and have been particularly associated with meningitis in neonates where infant-milk formulae has been epidemiologically linked to the disease. A study of the lipopolysaccharides produced by clinical isolates using chemical, 2D 1H and 13C NMR, and MS methods revealed that the O-polysaccharide produced by C. sakazakii (3290), a clinical strain from the Tennessee outbreak, was a branched polymer of repeating pentasaccharide units composed of 2-acetamido-2-deoxy-D-galactose, 3-(N-acetyl-L-alanylamido)-3-deoxy-D-quinovose, D-glucuronic acid, and D-glucose present in the molar ratio 1:1:1:2 and had the structure:The O-PS structure provides a unique specific structurally defined marker for the clinical tracking of this pathogen.

We examined the effect of growth inhibitory factor (GIF), also called metallothionein-III (MT-III), in brain damage using a stab wound model. The administration of 3 microM purified rat GIF (prGIF) provided significantly improved brain repair compared with controls, whereas the administration of 15 microM prGIF reduced brain repair compared with controls. To maintain the continuous effect of GIF, we generated an adenoviral vector encoding rat GIF and the myc epitope (AxCArGIFM) and administered an appropriate amount (1 x 10(8) pfu) of AxCArGIFM on the basis of the optimal dosage determined in a previous study on avulsion of the facial nerve. The administration of AxCArGIFM provided significantly improved histological and biochemical parameters of brain repair compared with controls administered AxCALacZ (adenovirus encoding bacterial beta-galactosidase gene as a reporter; 1 x 10(8) pfu). These results show that GIF can protect from brain damage in certain appropriate conditions in vivo and in vitro. The optimal dosage is very important for the treatment in vivo, particularly that for GIF. Our findings show the double-edged effects of GIF. MTs including MT-III are promising as therapeutic agents not only for tissue repair following acute brain injury, but also for some neurodegenerative diseases because they have multifunctional potential including anti-oxidation effects and may have some effect on neurogenesis.

Orf virus encodes a range of immuno-modulatory genes that interfere with host anti-virus immune and inflammatory effector mechanisms. The function of these reflects the pathogenesis of orf. The orf virus interferon resistance protein (OVIFNR) and virus IL-10 (vIL-10) inhibit interferon production and activity. In addition the vIL-10 suppresses inflammatory cytokine production by activated macrophages and keratinocytes. The virus GM-CSF inhibitory factor (GIF) is a novel virus protein that binds to and inhibits the biological activity of GM-CSF and IL-2. Together, these immuno-modulators target key effector mechanisms of host anti-virus immunity to allow time for virus replication in epidermal cells.

The intrinsic optical imaging technique has been widely applied for the visualization of functional maps in the sensory cortices of mammals. Many current studies refer this mapping in order to focus thereafter on particular features, at some particular locations: a fast and accurate mapping is therefore required. However, even during a successful experiment, the recorded raw data are usually contaminated by some kinds of noise that cannot necessarily be averaged out over the trials. An adequate image data analysis method has to be applied to extract signals closely related neural activities in response to presented stimuli. Thus far two different analysis methods could be adopted: the band-pass filtering and the GIF method [Yokoo T, Knight BW, Sirovich L. An optimization approach to signal extraction from noisy multivariate data. NeuroImage 2001:14;1309-26]. While the latter one is very efficient but requires the whole data in order to maximize the signal to noise ratio, the simple band-pass filtering technically reaches its limits very quickly. Here we propose another filtering method based on the polynomial subtraction of spatially smoothly modulated components. This simple method can visualize well-organized iso-orientation domains of the cat visual cortex with reliability similar to more sophisticated ones while allowing an online visualization of the clean data.

Human neuronal growth inhibitory factor (GIF) impairs the survival of cultured neurons and is deficient in the brains of Alzheimer's disease victims. We have isolated and sequenced analogous proteins from bovine and equine brain. By comparing their primary structures with those of human, mouse and rat GIFs, a consensus GIF sequence was obtained. Although this exhibits ca. 65% similarity with primary structures of mammalian metallothioneins (MTs), some significant differences are expected in the content of helix and turn secondary structures. In contrast to MTs, which usually bind 7 Zn(II) ions, human, bovine and equine GIFs contain 1-4 Cu(I) and 3-5 Zn(II) ions in species-specific ratios. The observed Cu(I) phosphorescence (lambda max, 550-590 nm; tau, 100 microseconds at 77 K) indicates the presence of the cuprous ion. Both bovine Cu1Cd5- and the equine Cu3Cd3-GIF derivatives (Cd replacing Zn) exhibit cadmium-dependent absorption and CD features between 220-260 nm characteristic of Cd-thiolate clusters similar to those in Cd-MTs.

Mice harboring a mutation in the gene encoding gastric intrinsic factor (Gif), a protein essential for the absorption of vitamin B12/cobalamin (Cbl), have potential as a model to explore the role of vitamins in infection. The levels of Cbl in the blood of Gif(tm1a/tm1a) mutant mice were influenced by the maternal genotype, with offspring born to heterozygous (high Cbl, F1) mothers exhibiting a significantly higher serum Cbl level than those born to homozygous (low Cbl, F2) equivalents. Low Cbl levels correlated with susceptibility to an infectious challenge with Salmonella enterica serovar Typhimurium or Citrobacter rodentium, and this susceptibility phenotype was moderated by Cbl administration. Transcriptional and metabolic profiling revealed that Cbl deficient mice exhibited a bioenergetic shift similar to a metabolic phenomenon commonly found in cancerous cells under hypoxic conditions known as the Warburg effect, with this metabolic effect being exacerbated further by infection. Our findings demonstrate a role for Cbl in bacterial infection, with potential general relevance to dietary deficiency and infection susceptibility.

Malnutrition continues to be a major public health problem in countries with weak infrastructures. In communities with a high prevalence of poor diet, malnourishment and infectious disease can impact vulnerable individuals such as pregnant women and children. Here, we describe a highly flexible murine model for monitoring maternal and environmental influences of vitamin B12 metabolism. We also demonstrate the potential importance of vitamin B12 in controlling susceptibility to bacterial pathogens such as C. rodentium and S Typhimurium. We postulate that this model, along with similarly vitamin deficient mice, could be used to further explore the mechanisms associated with micronutrients and susceptibility to diseases, thereby increasing our understanding of disease in the malnourished.

The purposes of this study were threefold: (1) to compare values obtained by three conventional radioimmunoassays for serum bone-gla-protein (BGP) in a population of normal women, (2) to study the relationship between serum BGP and bone mineral density (BMD) measured at four different skeletal sites (lumbar spine, proximal femur, proximal and ultradistal radius), and (3) to compare the results obtained by the three assays with conventional markers of bone turnover. Ninety-seven normal women (age range 25 to 75 years, mean +/- 1 SD = 54.3 +/- 10.9 years) were studied. Three independent assays were used to measure serum osteocalcin levels: a heterologous radioimmunoassay (RIA) (A) (Incstar Co., Stillwater, Minn.), a homologous RIA (B) (Nichols Institute, San Juan Capistrano, Calif.), and a two-site immunoradiometric assay (C) (Cis Biointernational, Gif-sur-Yvette, France). Mean +/- SD values of serum osteocalcin in the group as a whole were 4.05 +/- 1.37 microg/L by assay A, 6.03 +/- 2.90 microg/L by assay B, and 22.67 +/- 7.52 microg/L by assay C. Serum osteocalcin levels increased linearly with age; however, no correlation between serum BGP (whatever the assay used) and age was observed when only postmenopausal women were taken into account. When the effect of age was held constant by means of partial correlation analysis, only serum BGP levels measured by assays B and C were still inversely related with lumbar spine and ultradistal radius BMD; the latter assay was also weakly correlated with Ward's triangle BMD. After all the biochemical and clinical variables taken into consideration were introduced in a multiple regression equation, serum BGP still represented an important predictor of ultradistal radius and lumbar spine BMD only. Regarding relationships with other markers of bone turnover, the assay C in general showed the highest r values. In conclusion, our results indicate that commercially available BGP assays differ analytically and clinically; furthermore for the first time they show the existence of an inverse correlation between serum osteocalcin levels (which reflects bone turnover at the time of examination) and bone mass (which at a given time represents the balance of all previous metabolic events), after the influence of aging is excluded.

Assessment in vitro were carried out to study Toxo-GIF and IFN in lymphokines, LKs, produced by immune spleen cells with Toxoplasma lysate antigen (TLA). LKs were obtained from the cells after incubation for 24 to 48 h with TLA. They inhibited almost completely the intracellular multiplication of Toxoplasma (Tp) within homologous cells. Peak IFN activities were observed in the same LKs. The IFN was inactivated at pH 2, but did not lose its activity at 56 degrees C, suggesting that it might be type II or gamma IFN. Further evidence was presented of the coordinate production and properties of Toxo-GIF and IFN in the circulating blood of Tp-immune mice injected intraperitoneally with TLA or Poly I: C. The IFN was designated type II or gamma IFN. Its maximum activity appeared in Tp-immune mice 6 h after TLA or Poly I: C injection, while a maximum Toxo-GIF activity was observed in these mice 24 to 48 h after injection. Type I IFN activity was detected in the serum of normal mice injected with TLA or Poly I: C. Type I IFN differed markedly from type II IFN, because it was stable at pH 2 and unstable at 56 degrees C. Evidence was presented which emphasized the difference in activity in relation to the time of appearance between Toxo-GIF and IFN.

Growth inhibitory factor/metallothionein III (GIF/MT-III) is expressed specifically in brain, and neither mRNA nor protein is detected in other organs. This tissue-specific expression might be regulated by negative elements as well as positive elements, such as tissue-specific enhancers. To investigate the repression mechanisms of this gene in organs other than the brain, transfection experiments were performed by using various deletion mutants. Interestingly, a 25 x CTG repeat in the promoter region seemed to contribute to the repression activity. Moreover, the repression activity of this 25 x CTG repeat was also observed in various promoters and in a direction and position independent manner, indicating that this element could act as a silencer. However, no binding protein was detected by gel-shift and footprint analyses. These results strongly suggest that the CTG repeat functions as a negative element, and that this effect is caused by unknown mechanisms, rather than by interactions between specific cis-elements and specific trans-acting factors as reported previously. It is also possible that the CTG repeat functions as a general silencer in many genes.

During passive whole-body motion in the dark, the motion perceived by subjects may or may not be veridical. Either way, reflexive eye movements are typically compensatory for the perceived motion. However, studies are discovering that for certain motions, the perceived motion and eye movements are incompatible. The incompatibility has not been explained by basic differences in gain or time constants of decay. This paper uses three-dimensional modeling to investigate gondola centrifugation (with a tilting carriage) and off-vertical axis rotation. The first goal was to determine whether known differences between perceived motions and eye movements are true differences when all three-dimensional combinations of angular and linear components are considered. The second goal was to identify the likely areas of processing in which perceived motions match or differ from eye movements, whether in angular components, linear components and/or dynamics. The results were that perceived motions are more compatible with eye movements in three dimensions than the one-dimensional components indicate, and that they differ more in their linear than their angular components. In addition, while eye movements are consistent with linear filtering processes, perceived motion has dynamics that cannot be explained by basic differences in time constants, filtering, or standard GIF-resolution processes.

When Toxoplasma tachyzoites were inoculated into normal kidney cell monolayers, they multiplied in the cells and further causing cell rupture. However, when Toxoplasma immune lymphokines (LKs) was added to normal kidney cells, the intracellular multiplication of the tachyzoites was inhibited remarkably; particularly, in the case in which the tachyzoites were exposed to immune fresh serum prior to inoculation into the cells. Toxoplasma growth inhibitory factor (Toxo-GIF) in LKs, with a m.w. of 30,000 to 40,000 was found to be contained in LKs-II or MIF-I fraction after Sephadex G-100 gel filtration. LKs collected from Balb/c mice inhibited tachyzoite multiplication in ICR-JCL normal mice kidney cells, suggesting that LKs activity did not show mouse-strain specificity. In contrast, the addition of Toxoplasma immune serum to infected cells had no effect on the intracellular multiplication of Toxoplasma and also did not prevent cell-to-cell spread. However, when extracellular tachyzoites were exposed to Toxoplasma immune fresh or heat-inactivated immune serum, especially to the former, at 37 degrees C for 30 min, their active penetration into the kidney cells were extremely inhibited and their subsequent multiplication in the cells were almost totally inhibited within 24 h as compared with those exposed to normal serum.

Between January 1982 and December 1985, 355 fiberoptic pouchoscopies were performed in 123 patients with a continent ileostomy. These examinations have been reviewed to determine the effectiveness of the technique as a diagnostic and therapeutic tool. The Olympus GIF-XP pediatric endoscope was used after pouch lavage, and the afferent loop of ileum, the pouch, and (by retroflexion) the nipple valve were examined on each occasion. There were 63 males and 60 females, with a median age of 35 years (range, 16 to 71 years). The median length of follow-up after pouch construction was 36 months (range, 6 to 120 months), and an average of three examinations were performed per patient (range, 1 to 12). Of 127 examinations performed in asymptomatic patients, the pouch was normal in 117 cases, and there was mesh erosion into the pouch in 10 cases. The remaining 228 examinations were for symptoms that included pouchitis (56), difficulty in intubation (47), incontinence (35), follow-up of treated pouchitis (18), parastomal sepsis (22), blood in the stool (13), anemia (8), excess mucus discharge (6), valve prolapse (4), and purulent discharge from the stoma (1). Eighty-four examinations were normal; 144 revealed a likely cause for the symptoms and led to appropriate treatment, which in 45 patients was surgical. Fiberoptic endoscopy was therapeutic in 6 patients in whom it was used on 10 occasions to intubate a pouch with a slipped valve. Radiographic studies were seldom used, with pouchograms being carried out in 16 patients and fistulograms in 5. Only the fistulograms contributed to the assessment of each patient.(ABSTRACT TRUNCATED AT 250 WORDS)

Epidemiological parameters, such as age-dependent force of infection and average age at infection () were estimated for rubella, varicella, rotavirus A, respiratory syncytial virus, hepatitis A and parvovirus B19 infections for a non-immunized Brazilian community, using the same sera samples. The for the aforementioned diseases were 8.45 years (yr) [95% CI: (7.23, 9.48) yr], 3.90 yr [95% CI: (3.51, 4.28) yr], 1.03 yr [95% CI: (0.96, 1.09) yr], 1.58 yr [95% CI: (1.39, 1.79) yr], 7.17 yr [95% CI: (6.48, 7.80) yr] and 7.43 yr [95% CI: (5.68, 9.59) yr], respectively. The differences between average ages could be explained by factors such as differences in the effectiveness of the protection conferred to newborns by maternally derived antibodies, competition between virus species and age-dependent host susceptibility. Our seroprevalence data may illustrate a case of the above-mentioned mechanisms working together within the same population.

OBJECTIVE

Fluorochrome and immunohistochemical studies were performed on neonates with pontosubicular necrosis (PSN), aged 26 - 42 weeks of gestation (GW), compared with preterm and term controls aged from 10 GW to 3 months of age.

RESULTS

A fluorochrome study using a confocal microscope revealed that nuclear DNA changes occurred earlier than cytoplasm degeneration with diminished RNA orange-red fluorescence. These changes were restricted to the small immature neurons in the pons and subiculum with PSN. On the other hand, although glial fibrillary acidic protein-positive reactive astrocytes were not increased in number, growth inhibitory factor- (GIF) immunoreactive glia with vesicular large nuclei were increased in number within the gray matter of the pons, subiculum, and cerebral cortex in the PSN group. The nuclei of GIF-containing astrocytes became round and vesicular, nearly twice in size and increased in number. Thus, the neuronal death began at the nuclei of selective neurons in specific areas in PSN, although GIFcontaining astrocytes were increased in widespread areas.

CONCLUSIONS

These facts suggest that immature neurons in the pontine nuclei and subiculum are selectively vulnerable to some insults such as hypocarbia and hyperoxygenation, and PSN involves a possible apoptotic neuron death mechanism and a characteristic glial response.

The growth inhibitor factor (GIF) is a new member of the metallothionein family that is downregulated in Alzheimer's disease brain. Using a confocal laser scan microscope with polyclonal and monoclonal antibodies to GIF, and monoclonal antibodies to glial fibrillary acidic protein (GFAP) and MAP-2, we demonstrated that GIF immunoreactivity was expressed primarily in astrocytes and much less in neurons. In astrocytes of normal rat brain GIF immunoreactivity was detected mainly in the cell bodies, while GFAP immunoreactivity was detected mainly in the processes. GIF immunoreactivity was more strongly expressed in reactive astrocytes. These findings were confirmed with both polyclonal and monoclonal antibodies. Following stab wounds, a number of GIF-positive reactive astrocytes were detected around the wounds at 3 days postoperation. After 7 days GIF immunoreactivity was detected in cell bodies and processes of reactive astrocytes. The number of GIF-positive astrocytes and the intensity of the immunoreactivity remained elevated over the control levels at least through 28 days. These immunocytochemical findings correlated well with changes in GIF protein and mRNA levels. Not only changes in GIF protein and mRNA levels but also intracellular localization of GIF in normal rat brain and after stab wounds in rat brain were different from those of GFAP. These results support the concept that GIF plays an important role in the processing of reconstruction after brain damage.

BACKGROUND

The construction of evolutionary trees is one of the major problems in computational biology, mainly due to its complexity.

RESULTS

We present a new tree construction method that constructs a tree with minimum score for a given set of sequences, where the score is the amount of evolution measured in PAM distances. To do this, the problem of tree construction is reduced to the Traveling Salesman Problem (TSP). The input for the TSP algorithm are the pairwise distances of the sequences and the output is a circular tour through the optimal, unknown tree plus the minimum score of the tree. The circular order and the score can be used to construct the topology of the optimal tree. Our method can be used for any scoring function that correlates to the amount of changes along the branches of an evolutionary tree, for instance it could also be used for parsimony scores, but it cannot be used for least squares fit of distances. A TSP solution reduces the space of all possible trees to 2n. Using this order, we can guarantee that we reconstruct a correct evolutionary tree if the absolute value of the error for each distance measurement is smaller than f2.gif" BORDER="0">, where f3.gif" BORDER="0">is the length of the shortest edge in the tree. For data sets with large errors, a dynamic programming approach is used to reconstruct the tree. Finally simulations and experiments with real data are shown.

It was well known that beta-amyloid (Abeta) and tau protein play an important role in pathological procedure of Alzheimer's disease (AD), a senile dementia. The growth inhibitory factor (GIF, also named metallothionein-3, MT-3) had been demonstrated to inhibit the outgrowth of cortex neurons in the medium with extract of the AD patient brain. In our experiments, it was found that the neurons of cortex and the PC12 (pheochromocytoma) cells could be protected from the cytotoxicity of beta-amyloid 25-35 in presence of GIF and its domains. Additionally, GIF can scavenge the hydroxyl radical efficiently in CytC-VitC radical producing system and its alpha-domain shown more effective potentials than its beta-domain. The electron paramagnetic resonance spectra also show that the alpha-domain has more potential ability for eliminating reactive oxygen free radicals than its beta-domain. The results suggest that GIF could act as an efficient scavenger against free radicals in vitro and the alpha-domain in GIF molecule shows more potential in protecting against reactive oxygen species injury than the beta-domain.

We investigated growth inhibitory factor (GIF) mRNA expression within the rat facial nucleus with the aid of in situ hybridization. We found that GIF mRNA was expressed abundantly in the facial motoneurons of sham operated animals, and that this gene expression decreased after transection of the facial nerve. This decrease of GIF mRNA was first detected on the third day and was maintained for at least five weeks after transection of the nerve. Changes in c-jun, an immediate early gene, were also investigated with this model, and it was found that c-jun mRNA started to increase in the facial nucleus on the first day and that this increase was maintained for at least 5 weeks. These results suggest that the facial motoneurons, when their axons are transected, continuously respond to the injury and that GIF mRNA is actively suppressed to reduce the inhibition of neurite outgrowth in order to regenerate the axons.

Orf virus (ORFV), the type species of the family Parapoxviridae, encodes a protein (GIF) that binds and inhibits the ovine cytokines granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-2 (IL-2). There is no obvious sequence homology between the ORFV protein and any known mammalian GM-CSF- or IL-2-binding proteins. We demonstrate here that many of the biochemical properties of mammalian GM-CSF receptors that are required for efficient binding of GM-CSF are also critical to the GIF protein for binding to ovine GM-CSF (ovGM-CSF). Site-directed mutagenesis of the GIF protein demonstrated, first, the importance of disulfide bonds, and second, that a sequence motif (WDPWV), related to the WSXWS motif of the type 1 cytokine receptor superfamily, was necessary for biological activity. Finally, glycosylation of the GIF protein was also critical for binding to GM-CSF.

Humoral and cell-mediated immune responses of 36 captive foxes to two oral vaccines against rabies currently used for foxes in Europe were studied. The Street Alabama Dufferin (SAD) mutant Gif (SAG2) vaccine has been selected by double mutation from the SAD virus. The vaccinia recombinant virus (V-RG) expresses the rabies glycoprotein. Both vaccines induce similar humoral and cell-mediated responses after primary and secondary oral administration. We observed a typical anamnestic response, although of a limited duration, after the booster vaccination. Therefore, our results suggested that two successive oral vaccination campaigns should not significantly improve the immunisation of foxes. Lymphocyte in vitro proliferative response to the SAD antigen highlighted the presence in blood of a T-cell specific memory 6 months after vaccination. The synthesis of several vulpine cytokines was detected in peripheral blood mononuclear cells (PBMC) stimulated by SAD antigen via reverse transcription polymerase chain amplification. The data showed a concomitant expression of interleukin (IL)-4 and interferon-gamma in PBMC of vaccinated foxes. No change was detected in the level of IL-2, IL-10 and IL-12 synthesis, whereas the pro-inflammatory cytokine tumour necrosis factor-alpha seemed involved in the activation of naive T lymphocytes.

High-affinity binding was demonstrated between suppressor-T-cell-derived bioactive glycosylation-inhibiting factor (GIF) and helper T hybridomas and natural killer cell line cells. Inactive GIF present in cytosol of suppressor T cells and Escherichia coli-derived recombinant human GIF (rhGIF) failed to bind to these cells. However, affinity of rhGIF for the target cells was generated by replacement of Cys-57 in the sequence with Ala or of Asn-106 with Ser or binding of 5-thio-2-nitrobenzoic acid to Cys-60 in the molecule. Such mutations and the chemical modification of rhGIF synergistically increased the affinity of GIF molecules for the target cells. The results indicated that receptors on the target cells recognize conformational structures of bioactive GIF. Equilibrium dissociation constant (Kd) of the specific binding between bioactive rGIF derivatives and high-affinity receptors was 10-100 pM. Receptors for bioactive GIF derivatives were detected on Th1 and Th2 T helper clones and natural killer NK1.1(+) cells in normal spleen but not on naive T or B cells. Neither the inactive rGIF nor bioactive rGIF derivatives bound to macrophage and monocyte lines or induced macrophages for tumor necrosis factor alpha production.

A human T-T hybridoma clone with helper cell phenotype was established from fusions between a HPRT- variant of the human T-cell lymphoma Molt4, and PPD-activated normal human T-lymphocytes. The hybrid clone (MP-6) was characterized with regard to expression of markers and lymphokine secretion. The T-T hybridoma was positive for Leu 3a, and thus of T-helper cell lineage. The transferrin receptor (T-9) and the interleukin 2 (IL-2) receptor (Tac) were also expressed as judged by immunofluorescence analysis using monoclonal antibodies. The hybridoma produces B-cell stimulatory factor (BSF) with proliferation and maturation activities, growth inhibitory factor (GIF), leukocyte migration inhibitory factor (LIF) constitutively under serum free conditions, but no detectable interferons (IFN-alpha, IFN-beta, IFN-delta), nor interleukin 2 (IL-2). Weak interleukin 1 (IL-1)-like activity was found. The B-cell stimulatory factor (BSF) induced solid phase-anti-mu triggered resting B-cells obtained from human spleen, tonsil or peripheral blood to proliferate and to secrete IgM and IgG. Without anti-mu triggering the BSF had no proliferation inducing effect. The BSF was characterized and partially purified using ammonium sulphate precipitation, Blue-Sepharose, HPLC hydrophobic interaction and HPLC gel filtration chromatography. The BSF was heat labile at 56 degrees C and was present in two forms, one with high and one with intermediate hydrophobicity. The more hydrophobic form of BSF has a molecular weight of 12K-14K. Kinetic studies of the lymphokine secretion revealed that BSF was produced in detectable amounts in low density (0, 2 X 10(6) cells/ml) 18-24 h cultures. In 48 h to 72 h cultures there was a significant influence of growth inhibitory activities (GIF) produced. GIF, with an apparent MW of 90K could be absorbed out on certain tumor cell lines or on Blue-Sepharose. Further absorption analysis of BSF activities show that anti-mu triggered B-cells but neither resting B-cells nor T-cells could absorb BSF activity.

OBJECTIVE

Mesothelin (SMRP) is regarded as a biomarker of malignant pleural mesothelioma (MPM). Herein, we analyzed the contribution of SMRP detection in pleural effusion and in serum to the diagnosis of MPM with non-positive cytology.

METHODS

The present study included 52 cases of MPM, 43 of pleural benign lesions and 25 of non-MPM pleural metastases. SMRP was measured by MesoMark ELISA (Cis-Bio International Gif/Yvette; France).

RESULTS

In non-positive cytology, effusion-SMRP showed higher diagnostic performance than serum-SMRP. We found 38 out of 52 (73.1%) cases of non-positive cytology MPM, out of which 27 (71.0%) were positive for effusion-SMRP (cut-off=12.70 nM) and 18 (47.4%) for serum-SMRP (cut-off=1.08 nM). When cytology, effusion- and serum-SMRP were used in combination, an overall sensitivity in detection of MPM of 78.9% was achieved. The same sensitivity was obtained by combining cytology with effusion-SMRP alone, whereas the combination of serum-SMRP with cytology led to a sensitivity of 61.5%.

CONCLUSIONS

Detection of both effusion- and serum-SMRP can contribute to improve the diagnosis of MPM with non-positive cytology. However, the analysis of SMRP in effusion makes it unnecessary to test SMRP in the serum.

Steroid hormones, especially glucocorticoids, exert physiologic effects on dopaminergic neurotransmission and have been implicated in several dopamine-mediated neuropsychiatric conditions. D(2) dopamine receptor gene expression is regulated by the zinc finger-type nuclear protein GDNF-inducible transcription factor (GIF). In this study, we sought to investigate if steroids could regulate transcription of the GIF gene itself. Transient co-transfection of the D(2) expressing neuroblastoma cell line NB41A3 with GIF promoter-luciferase constructs along with expression vectors for steroid hormone receptors showed that activation of glucocorticoid receptors but not estrogen receptors up-regulates transcription from the GIF promoter 5.0-fold. Progesterone receptors, which share the same consensus DNA recognition sequence as glucocorticoid receptors, also activated the GIF promoter. Serial 5'-deletion mutants of the GIF gene upstream region localized the glucocorticoid-responsive segment between nucleotides -128 and -66 relative to the transcription start site. This region contains a putative glucocorticoid-responsive element/progesterone-responsive element (GRE/PRE). Additionally, this fragment of the GIF gene 5'-upstream region activated the heterologous herpes simplex virus thymidine kinase (TK) promoter, which is known to be glucocorticoid and progesterone responsive. Furthermore, glucocorticoid receptor activation up-regulated endogenous GIF gene mRNA expression in NB41A3 cells. These observations demonstrate a molecular basis for glucocorticoid and progesterone-induced up-regulation of GIF gene transcription and provide a mechanism for the modulation of dopamine-mediated behaviors by these hormones.