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We have established lines and clones of helper T cells specific for the antigen fowl gamma globulin (FGG) and maintained these cells in continuous proliferation in vitro for up to 8 months. During this period the specific helper activity was assessed in vitro in T cell-dependent anti-hapten responses. These pure populations of FGG-specific helper T cells provide help in the absence of other T cells for the induction of B cells to produce specific antibody of different isotypes and idiotypes. The same clones of specific helper T cells can provide help for a T cell-dependent antibody response against azobenzene arsonate (ARS), dinitrophenol (DNP) or phosphorylcholine (PC). With the exception of one clone all FGG-specific helper T cells (TFGG) promoted anti-hapten responses only when the hapten was covalently bound to the specific carrier. Help was only provided when T cells and T-depleted spleen cells were H-2 compatible to the left of the I-B region. [ABSTRACT FROM AUTHOR]

Plasma levels of tumor necrosis factor-α (TNFα), interleukin-1 (IL-1),and interleukin-6 (IL-6) were monitored after intravenous administration of Escherichia coli endotoxin with or without ibuprofen pretreatment to healthy volunteers. Intravenous endotoxin (n = 7) resulted in elevated plasma TNFα concentrations with maximal levelsat 90 min (369 ± 44 pg/ml, P < .001 vs. saline controls, n = 7). The rise in TNF-α was followed by a rise in plasma IL-6 (27 ± 12.8 ng/ml), peaking 30–90 min thereafter. Pretreatment with ibuprofen (n = 6) caused a significant augmentation and temporal shift in cytokine elaboration with maximal TNFα levels(627 ± 136 pg/ml) at 120 min and IL-6 peaks (113 ± 66 ng/ml) at 180 min. In ibuprofen-treated volunteers, the additional increase in TNFα was paralleled by increased levels of circulating elastase. In vitro experiments suggest a causal relationship between these events. Thus, the cyclooxygenaseinhibitor ibuprofen blunts the clinical response to endotoxin but augments circulating cytokine levels and leukocyte degranulation. [ABSTRACT FROM PUBLISHER]

Background: Cancer patients with chemotherapy-induced peripheral neuropathy (CIPN) have sensory and motor deficits leading to inappropriate proprioceptive feedback, impaired postural control, and fall risk. Balance training with computerized force platforms has been successfully used in rehabilitation of balance disturbances, but programs specifically developed for CIPN patients are lacking., Aim: This pilot study evaluated a rehabilitation protocol exclusively based on visual computer-feedback balance training (VCFBT) to improve balance in patients with CIPN., Design: Open-label, non-randomized pilot study, 4-week intervention with pre- vs. post-treatment evaluation., Setting: Outpatients of the Rehabilitation Institute of the Salvatore Maugeri Foundation, in Genoa, Italy., Population: Seven out-patients with clinical-instrumental diagnosis of CIPN., Methods: At admission, patients were administered the Berg Balance Scale (BBS) and underwent static-dynamic posturography using a computerized force platform to objectively quantify their balance impairment. Their performance was compared to values of a normal age-matched population. Patients then underwent 4 weeks of VCFBT (three 60-minute sessions/week). At discharge, BBS and posturography were repeated and the results compared with those at admission., Results: A significant pre- vs. post-treatment improvement was found in balance as measured by static-dynamic posturography (P=0.004) and BBS (P<0.002)., Conclusions: Despite caution needed for the low sample size, this pilot study has shown preliminary evidence that intensive rehabilitation, based on VCFBT can produce a significant improvement in balance outcomes., Clinical Rehabilitation Impact: To our knowledge, this is the first report in CIPN patients of a rehabilitation program based exclusively on VCFBT.

It is not known whether postsurgery systemic inflammation and plasma amino acid abnormalities are still present during rehabilitation of individuals after elective hip arthroplasty (EHA). Sixty subjects (36 females; age 66.58 ± 8.37 years) were randomized to receive 14-day oral EAAs (8 g/day) or a placebo (maltodextrin). At admission to and discharge from the rehabilitation center, serum C-reactive protein (CRP) and venous plasma amino acid concentrations were determined. Post-EHA hip function was evaluated by Harris hip score (HHS) test. Ten matched healthy subjects served as controls. At baseline, all patients had high CRP levels, considerable reduction in several amino acids, and severely reduced hip function (HHS 40.78 ± 2.70 scores). After treatment, inflammation decreased both in the EAA group and in the placebo group. Only EAA patients significantly improved their levels of glycine, alanine, tyrosine, and total amino acids. In addition, they enhanced the rate of hip function recovery (HHS) (from baseline 41.8 ± 1.15 to 76.37 ± 6.6 versus baseline 39.78 ± 4.89 to 70.0 ± 7.1 in placebo one; p = 0.006). The study documents the persistence of inflammation and plasma amino acid abnormalities in post-EHA rehabilitation phase. EAAs enhance hip function retrieval and improve plasma amino acid abnormalities.

Background: Recent data suggest that propranolol is an effective treatment for infantile hemangiomas (IHs). Data on the optimal dose, duration of therapy, and predictors of response are currently lacking., Objective: To assess the clinical response to and predictors of propranolol use in the treatment of IH., Methods: Retrospective cohort study of 44 patients. Two independent assessors evaluated improvement by comparing serial digital photographs using a 100 mm visual analogue scale (VAS), where 5 mm change represented 10% change in the size or appearance of the IH., Results: Propranolol was started at a mean age of 7.8 (SD 8.21) months and was used for 7.3 (SD 4.8) months before weaning. The mean percent improvement compared to baseline (as measured by the VAS) was 78% (SD 23%). Minor adverse events were noted in 32% of patients. The most significant predictor of regrowth after weaning was a IH > 5 cm in size (p  =  .017)., Conclusions: Propranolol is effective in IH, but the side effects and the possibility of regrowth should be considered.

Primitive myxoid mesenchymal tumor of infancy is a recently recognized entity that has been added to the differential diagnosis of myxoid tumors of the soft tissue. Few cases have been reported of this entity in the literature, but none presenting in a preterm infant. We present the case and clinical course of a preterm boy with a primitive myxoid mesenchymal tumor of infancy that occurred following excision of a congenital juvenile xanthogranuloma., (© 2010 Wiley Periodicals, Inc.)

We evaluated alterations of balance by stabilometry in patients with amnestic mild cognitive impairment (aMCI) and with mild-moderate Alzheimer's disease (AD). Fifteen patients with aMCI and 15 with mild AD were recruited according to the current diagnostic criteria. Fifteen healthy subjects of the same age range were recruited as controls. Stabilometry was carried out using a commercial 4 load cell platform. Statistical analysis of between group differences was performed using one-way analysis of variance for parametric data and Kruskal-Wallis tests for non-parametric data. Spearman correlation coefficients were used to investigate the association between cognitive test scores and stabilometric data. All stabilometry measures were significantly altered in mild AD patients compared to normal controls. Antero-posterior sway was found to be the most sensitive parameter, since it correlated with the ADAS-cog orientation subscale in AD patients, and also discriminated between aMCI and normal controls. Our study shows that impairment in balance is a feature not only of AD, but also of aMCI. The alterations found suggest that a progressive failure of the vestibular system, possibly linked to reduced hippocampal performance, may be responsible for such a feature. Further research must be focused on studying the predictive value of stabilometry in the conversion of aMCI.

IL-5 production in vivo plays a unique role in the production, activation, and localization of eosinophils in a variety of allergic conditions. The current paradigm suggests that allergen-specific Th2 cells are the main source for the IL-5 production. The experiments outlined in this work, however, suggest that in vivo production of IL-5 by NK cells can separately influence eosinophil-associated inflammatory responses. Specifically, a mouse model of allergic inflammation was used in which C57BL/6 mice were immunized and challenged with a short ragweed Ag extract, known to induce a selective eosinophilia within the peritoneal cavity. Peritoneal lavage fluids from these mice also contained increased numbers of T cells and NK cells, as well as significantly elevated levels of IL-4, IL-5, and IFN-gamma. Flow-cytometric analysis of cytokine-producing cells in peritoneal lavage fluid revealed increased numbers of IL-5-producing cells in both T cell and NK cell populations following allergen exposure. Depletion of NK cells by treatment with NK1.1 Abs selectively reduced the number of infiltrating eosinophils by more than 50%. Moreover, the inhibition of the infiltration of eosinophils was accompanied by a complete loss of IL-5-producing NK cells and significantly reduced levels of peritoneal lavage fluid IL-5, whereas the number of IL-5-producing T cells was not affected. Thus, the results presented in this study provide clear evidence for a novel immunoregulatory function of NK cells in vivo, promoting allergen-induced eosinophilic inflammatory responses by the production of IL-5.

The chemical derivatization of biologically active microbial metabolites continues to be a promising approach to the identification of new drugs. We recently synthesized the novel antiproliferative compound SDZ 281-977, 5-[2-(2,5-dimethoxy-phenyl)ethyl]-2-hydroxy-benzoic acid methylester, a derivative of the EGF receptor tyrosine kinase inhibitor lavendustin A. Here we report on our studies of the anticancer efficacy and the mode of action of SDZ 281-977. The growth of both the human pancreatic tumor cells MIA PaCa-2 and the human vulvar carcinoma cells A431 was inhibited in the low micromolar range. Tumors from these cells were induced in nude mice and were shown to respond to orally or intravenously administered SDZ 281-977. In contrast, no antitumor effect was detected in rats bearing dimethylbenzanthracene-induced mammary tumors. Studies in mice indicated that SDZ 281-977 was neither immunosuppressive nor hematosuppressive at doses effectively inhibiting tumor growth. Surprisingly, the mode of action of SDZ 281-977 apparently does not involve inhibition of EGF receptor tryosine kinase, because, in contrast to lavendustin A, SDZ 281-977 failed to inhibit this enzyme in a cell-free assay. The mechanism of the antiproliferative effect can be explained on a cellular level by the ability of the compound to arrest cells in mitosis. SDZ 281-977 is thus the first example of an antimitotic agent derived from the potent tyrosine kinase inhibitor lavendustin A. The therapeutic potential of SDZ 281-977 is enhanced by the fact that it is not subject to multidrug resistance, because tumor cells expressing the multidrug resistance phenotype were as sensitive to SDZ 281-977 as their nonresistant counterparts. In conclusion, SDZ 281-977 represents a novel lavendustin A derivative with potent antiproliferative properties in vitro and in vivo that may be explained on the basis of its antimitotic effects. SDZ 281-977 may be a candidate drug for the treatment of selected cancers, including those expressing the multidrug resistance phenotype.

The active partial structure of the potent tyrosine kinase inhibitor lavendustin A was derivatized in the search for novel agents against cellular proliferation. The antiproliferative potential of the new derivatives was determined using the human keratinocyte cell line HaCaT as the primary test system. Whereas the lavendustin A partial structure is ineffective in inhibiting cell proliferation, esterification of its carboxylic acid function leads to measurable antiproliferative activity. Additional O-methylation of the 2,5-dihydroxyphenyl moiety yields activity in the micromolar range. Further substantial increases in activity are achieved by replacing the nitrogen with oxygen and carbon within the 2,5-dimethoxyphenyl series (but not within the 2,5-dihydroxyphenyl analogs) leading to 5-[2-(2,5-dimethoxyphenyl) ethyl]-2-hydroxybenzoic acid methyl ester (13) as the most potent analog identified to date. These increases in antiproliferative activity are paralleled, however, by the disappearance of activity against the epidermal growth factor receptor-associated tyrosine kinase, suggesting another mechanism of action.

In the absence of the homologous antigen, an anti-phosphorylcholine (PC) plaque-forming cell (PFC) response has been induced in vitro which is restricted to the TEPC 15 idiotype (T15). Anti-T15 antibodies were used to focus either the keyhole limpet hemocyanin (KLH) or the fowl gamma-globulin (FGG) carrier molecules on the membrane of B cells carrying the T15 idiotype on their immunoglobulin receptors; thereafter, these cells were allowed to cooperate in vitro for 5 days with T cells primed to the appropriate carrier molecule. A response, virtually 100% T15+, could be induced both in normal BALB/c and in T15 neonatally suppressed mice which had lost the T15+ clonal dominance. The magnitude of this response is comparable to that obtained in the presence of the PC antigen. The role of the membrane-bound immunoglobulin receptor (sIg) in the expression of different anti-PC clones was also investigated. We have focused either the KLH or the FGG carrier molecules on the membrane of B cells via anti-H-2 antibodies and then cultured these cells for 5 days with the appropriate carrier-specific T cells. Under these conditions, B cells are activated in the absence of interaction at the sIg. The idiotypic profile of the anti-PC PFC obtained with the anti-H-2-mediated activation was then compared with the profile of the anti-PC response obtained in the presence of PC antigen. Since similar idiotypic profiles were obtained in both cases, it can be excluded that sIg plays a direct role in favoring the expression of T15+ over T15- anti-PC clones.

Using the same anti-TNP hybridoma supernatant pool as IgE antibody source (2 micrograms/ml), several methods were compared for their sensitivity in IgE detection and convenience for screening purposes. Using rat basophilic leukemia (RBL) cells as specific receptor cells, one out of two rosetting methods with TNP-sheep red blood cells allowed the detection of 0.5 ng IgE/ml (50 pg/assay) while the other was much less sensitive (60 ng/ml). More convenient for screening was an ELISA method performed on cells, in which the IgE bound to the RBL cell surface could be detected either by enzyme-anti-Ig or by enzyme-antigen conjugates with a similar, albeit low, sensitivity of 10 ng IgE/ml (500 pg/assay). Using the same antibody and cell sources, a very convenient and more sensitive method for screening purposes was found to be the measurement of antigen-induced basophil granule beta-N-acetylglucosaminidase release by IgE sensitized RBL cells: 0.5 ng IgE/ml and 50 pg/assay. For the same anti-TNP IgE source, this compares with a detection limit by ELISA of 0.2 ng IgE/ml (10 pg/assay) and by passive cutaneous anaphylaxis in rats of 2 ng IgE/ml (100 pg/assay).

An aqueous extract of Panax Ginseng C.A. Meyer (G.S.) was prepared by boiling crushed G.S. roots in water. The extract obtained was adjusted to 125 mg G.S. per ml and was administered orally to mice for 5 to 6 days at the daily dose of 10, 50 and 250 mg G.S. per kg or was added to cultures of mouse spleen cells at concentrations varying between 0.25 and 8 mg G.S. per ml. The average total ginsenoside content of the G.S. roots used was determined by HPLC analysis and found to be 0.58% (w/w). Treated mice responded with enhanced antibody formation to either a primary or a secondary challenge with sheep red cells. The effects were dose-dependent. At the highest dose regimen, the primary IgM response was increased by 50% and the secondary IgG and IgM responses were increased by 50 and 100%, respectively. An even more pronounced effect was obtained with natural killer cell activity which was enhanced between 44 and 150% depending on the effector-to-target cell ratios used in the assay. In vitro, G.S. showed two main effects, an inhibition of stimulated and spontaneous lymphocyte proliferation at high, but not cytotoxic concentrations and an enhancement of interferon production particularly in non-stimulated spleen cells. The immunostimulating effects obtained in vivo are in agreement with the stimulation of interferon production observed in vitro. The inhibition of lymphocyte proliferation, however, cannot be reconciled with the immunostimulatory action of G.S. observed in vivo.

Mouse T lymphocytes cultured in the presence of Vicia villosa lectin, usually considered to be non-mitogenic, proliferate and produce interleukin-2. V. villosa-induced proliferation does not correlate with the cell ability to adhere to V. villosa-coated dishes, this is evident from the fact that either V. villosa-adherent or -nonadherent cells are equally sensitive to the mitogenic effect of the lectin. Furthermore, the sugar N-acetyl-d-galactosamine, which is reported to be highly specific for V. villosa, does not reduce the lectin-induced proliferation. These data indicate that there is a difference in the carbohydrate moieties required for the adherence to V. villosa and its mitogenic effect. These results are discussed on the light of the current use of V. villosa lectin as marker for some T cell subpopulations.

Bifunctional reagents have been prepared which permit extensive coupling of haptens to antibodies with retention of antibody-binding capacity. This modification procedure increases the sensitivity with which cell-surface antigens can be labeled for fluorescent or electron microscopy by the hapten-sandwich method to a level comparable to or greater than may usually be achieved by conventional indirect techniques. With two different haptens, and amplifying with anti-hapten antibodies bearing fluorescent or other markers, different antigens can be distinguished readily on a single cell or on separate cells. Mouse alloantigens detected only with difficulty by conventional fluorescence can be discerned clearly and with high specificity. The reagent, either methyl hydroxybenzimidate (HB) or methyl 3,5-dihydroxybenzimidate (DHB), is first reacted with a diazonium phenyl hapten. Since the resulting azo-derivative retains the imido ester function, the reaction solution can be used directly to amidinate antibodies. Both HB and DHB are easily prepared and may be stored for use as needed. With these reagents, the hapten-sandwich procedure may be applied to proteins other than immunoglobulins, e.g., hormones and mitogens. In addition to labeling antigens for fluorescent or electron microscopy, haptenamidinated antibodies or other ligands may be used in conjunction with appropriately modified antihapten amplifiers for a variety of purposes in cell biology, such as radioisotope studies, selective cell killing or suppression, and isolation of membrane antigens.

In order to investigate the molecular mechanism of biological action of the drug cyclosporin A (CsA), we have studied its effect on eukaryotic RNA polymerase activities in two systems: (a) in vitro transcription; (b) injection into frog oocytes. The electron microscopic transcription R-loop method [1] has been used to assay for specific transcription of an immunoglobulin gene clone in HeLa cell lysates. In this system, CsA inhibits efficiently the transcription of immunoglobulin genes. Dilution experiments show that RNA polymerase II activity is inhibited by nanomolar concentrations (i.e. amounts comparable to the ones needed for the inhibition of lymphocyte stimulation), whereas polymerase III is inhibited at micromolar concentrations. In vitro transcription with E. coli RNA polymerase is not inhibited. Parallel experiments with the mushroom toxin alpha-amanitin have shown that the eukaryotic RNA polymerase II displays similar sensitivity towards the two cyclic peptides CsA and alpha-amanitin in vitro. When CsA is injected into frog oocytes, together with a cloned Xenopus laevis U1 small nuclear RNA gene, transcription of this gene by endogenous RNA polymerase II is inhibited by 60%. RNA polymerase III and RNA polymerase I transcription is not inhibited in the oocyte. Various possibilities for the selective action of CsA on stimulated lymphocytes are discussed.

This article describes the construction and establishment of a double congenic nude, beige C57BL/6 (B6 nu, bg) mouse strain. The mice do not show higher fragility than C57BL/6 nude mice and the double congenic strain can be maintained under conventional mouse housing conditions. Although the B6 nu, bg display a very low natural killer activity which cannot be enhanced by an interferon inducer (poly(I-C], they lack responsiveness to a T cell mitogen (concanavalin A); and they also show extremely low responsiveness to a B cell mitogen (0128: B12 Escherichia coli lipopolysaccharide) probably as a result of combined effects of the beige and nude genes in the C57BL/6 genetic context.

T10B9.1A-31 is an immunoglobulin Mk (lgMk), CD3b class, murine monoclonal antibody. It is not itself mitogenic, but it partially blocks mitogenesis produced by concanavilin-A, phytohemagglutinin, and a soluble antigen cocktail and is lytic for human peripheral blood T lymphocytes (PB-T) in the presence of fresh autologous human complement and rabbit complement. It reacts with a monomorphic epitope found on all mature PB-Ts that modulates in vitro and in vivo with the CD3/T-cell antigen receptor (TCR) complex, but unlike OKT3, which reacts with CD3 proteins, T10B9.1A-31 reacts with an epitope of the TCR alpha/beta heterodimer. Immunoprecipitation of HPB-ALL with T10B9.1A-31 or OKT3 revealed that T10B9.1A-31 does not react with the 20-to 26-kd polypeptides that compose CD3, and T10B9.1A-31 failed to bind to the PEER cell, which is CD3 gamma/delta positive, TCR alpha/beta negative. Phase 1 clinical trials demonstrated that T10B9.1A-31 caused a rapid decrease in PB-Ts and no toxic effects other than fever, chills, rigors, and transient hypotension. These side effects were absent in methylprednisolone (MP)-pretreated patients. Phase II studies revealed that T10B9.1A-31 reversed steroid refractory acute rejection in three of five renal allografts treated with cyclosporine (CyA) and prednisone (P) maintenance immunosuppression, reversed steroid and polyclonal antisera refractory rejection in three cardiac allograft recipients receiving CyA/P, and attenuated acute rejection in three of three renal patients receiving baseline azathioprine/prednisone (AZA/P) when used as primary therapy. Rerejection was not seen when patients received CyA baseline immunosuppression but was seen in patients maintained on AZA/P OKT3 was efficacious in treating rejection following T10B9.1A-31 therapy. Side effects of T10B9.1A-31 appears to reverse acute rejection crisis effectively in renal and cardiac transplantation with a low incidence of rerejection in CyA-maintained patients and with fewer, milder side effects. Sequential therapy with OKT3 is possible because OKT3 and T10B9.1A-31 are of different isotypes and idiotypes.

Catecholamine and indoleamine neurotransmitters, together with some of their precursors and metabolites, were determined using HPLC in three brain and two spinal cord regions of Lewis rats with chronic relapsing allergic encephalomyelitis and of control rats injected with complete Freund's adjuvant. Three attacks and two recovery phases were investigated. Changes are found mainly in the spinal cord. In the lumbosacral region both 5-hydroxytryptamine and noradrenaline are reduced during the entire course of the disease, whereas in the craniothoracal region 5-hydroxytryptamine is unchanged and only noradrenaline is reduced during the attacks, returning to normal during the first recovery. The precursors tyrosine and tryptophan are greatly elevated during the first two attacks in both regions. The 5-hydroxytryptamine turnover marker 5-hydroxyindoleacetic acid is increased in the first attack in both regions, then it decreases in the later stages, indicating destruction of nerve fibers. On the fourth and seventh days after inoculation values are generally not significantly different from controls in all regions. The possible correlation of neurochemical results with neurological signs is discussed.