Macrophages that have ingested apoptotic cells in vitro inhibit proinflammatory cytokine production through autocrine/paracrine mechanisms involving TGF-beta, PGE2, and PAF

Apoptosis in vivo is followed almost inevitably by rapid uptake into adjacent phagocytic cells, a critical process in tissue remodeling, regulation of the immune response, or resolution of inflammation. Phagocytosis of apoptotic cells by macrophages has been suggested to be a quiet process that does not lead to production of inflammatory mediators. Here we show that phagocytosis of apoptotic neutrophils (in contrast to immunoglobulin G-opsonized apoptotic cells) actively inhibited the production of interleukin (IL)-1beta, IL-8, IL-10, granulocyte macrophage colony-stimulating factor, and tumor necrosis factor-alpha, as well as leukotriene C4 and thromboxane B2, by human monocyte-derived macrophages. In contrast, production of transforming growth factor (TGF)-beta1, prostaglandin E2, and platelet-activating factor (PAF) was increased. The latter appeared to be involved in the inhibition of proinflammatory cytokine production because addition of exogenous TGF-beta1, prostaglandin E2, or PAF resulted in inhibition of lipopolysaccharide-stimulated cytokine production. Furthermore, anti-TGF-beta antibody, indomethacin, or PAF receptor antagonists restored cytokine production in lipopolysaccharide-stimulated macrophages that had phagocytosed apoptotic cells. These results suggest that binding and/or phagocytosis of apoptotic cells induces active antiinflammatory or suppressive properties in human macrophages. Therefore, it is likely that resolution of inflammation depends not only on the removal of apoptotic cells but on active suppression of inflammatory mediator production. Disorders in either could result in chronic inflammatory diseases.     

Lipopolysaccharide induction of MARCKS-related protein and cytokine secretion are differentially impaired in microglia from LPS-nonresponsive (C3H/HeJ) mice

Many events involved in activation of microglia and leukocytes by lipopolysaccharide (LPS) are mediated by protein kinase C (PKC), and we have recently demonstrated that a major PKC substrate, MARCKS-related protein (MRP), is selectively induced by LPS in murine microglia. In microglia from LPS-nonresponsive (C3H/HeJ) mice, induction of MRP and secretion of CSF-1 required much higher LPS concentrations (> or = 100 ng/ml) than in normal (C3H/OuJ) microglia (< or = 10 ng/ml). By contrast, TNF alpha production was not significantly increased in C3H/HeJ microglia even at 1 microgram LPS/ml. Microglia expressed PKC isoforms alpha, beta, delta, and zeta (but not gamma and epsilon); PKC isoform levels were similar in both normal and C3H/HeJ microglia and no significant change in response to LPS was noted. Our results indicate that LPS alters PKC substrate (rather than kinase) expression, and that the Lpsd mutation in C3H/HeJ mice differentially affects regulation of several gene products implicated in microglial function.     

The effect of simian immunodeficiency virus infection in vitro and in vivo on the cytokine production of isolated microglia and peripheral macrophages from rhesus monkey

Microglia are the major target for human immunodeficiency virus (HIV) infection within the central nervous system. Because only a few cells are productively infected, it has been suggested that an aberrant cytokine production by this cell population may be an indirect mechanism leading to the development of neurological disorders in HIV-infected patients. Therefore we decided to study the secretion pattern of several interleukins (IL) by microglial cells and peripheral blood macrophages isolated from uninfected and simian immunodeficiency virus (SIV)-infected Rhesus monkeys. We found that uninfected, unstimulated primate microglia produce more IL-6 and less TNF alpha than peripheral blood macrophages, but generate comparable levels of IL-1 beta and IL-8. After infection with SIV in vitro, synthesis of all cytokines tested is increased compared to uninfected cultures and to peripheral blood macrophages. Microglia isolated from infected animals produce more IL-8 and TNF alpha than the uninfected cultures and display a strongly increased capacity to secrete TNF alpha upon stimulation with lipopolysaccharide. In addition, production of IL-6 by in vivo-infected microglia increases with time in culture to very high levels despite the fact that only a few cells contained replicating virus. These findings clearly show that the cytokine production of microglia is impaired after SIV infection both in vitro and in vivo and that a low level of viral replication is sufficient for these alterations to occur. In conclusion, the results of this study further support a possible role of cytokines in the pathogenesis of neuro-AIDS.     

Modulation of T cell cytokine profiles and peptide-MHC complex availability in vivo by delivery to scavenger receptors via antigen maleylation

We have previously shown that conversion of proteins to scavenger receptor (SR) ligands by maleylation increases their immunogenicity. We now show that maleyl-Ag-immune spleen cells make relatively more IFN-gamma and less IL-4 or IL-10 than native Ag-immune cells. This is also reflected in the IgG1:IgG2a ratios in Abs generated in vivo. SR engagement on macrophages does not alter their surface levels of the adhesive/costimulatory molecules CD11a/CD18, CD11b/CD18, CD24, CD54, or CD40, nor does it enhance their ability to support anti-CD3-driven proliferation of naive T cells in vitro. Costimulatory molecules implicated in differential Th1/Th2 commitment--CD80, CD86, and IL-12--are not inducible by SR ligation. In addition to macrophages and dendritic cells, B cells also show receptor-mediated uptake and enhanced presentation of maleyl-Ags. Using a monoclonal T cell line to detect peptide-MHC complexes expressed on spleen cells in Ag-injected mice, we find that higher levels of these complexes are generated in vivo from maleyl-proteins and they persist longer than those generated from the native protein. Together, these data suggest that in certain situations, the levels of cognate ligand available and/or the time course of their availability may play a major role in determining the cytokine profiles of the responding T cells in addition to the costimulatory signals implicated so far.     

Pathogenic mechanisms in the rheumatoid nodule: comparison of proinflammatory cytokine production and cell adhesion molecule expression in rheumatoid nodules and synovial membranes from the same patient

OBJECTIVE: To investigate the production of proinflammatory cytokines and expression of cell adhesion molecules in the rheumatoid nodule. METHODS: Cytokine content (tumor necrosis factor alpha [TNFalpha], interleukin-1beta [IL-1beta], and IL-1 receptor antagonist [IL-1Ra]), at the messenger RNA (mRNA) and protein levels, and cell adhesion molecule expression were studied in 16 rheumatoid nodules and 6 synovial membranes. RESULTS: Macrophages in the rheumatoid nodules contained TNFalpha, IL-1beta, and IL-1Ra mRNA and protein, particularly in perivascular cells of the stroma and in the palisading layer. All cell adhesion molecules studied were expressed in both the rheumatoid nodules and synovial membranes, with increased expression of E-selectin in the rheumatoid nodule compared with the synovial membrane, and with the absence of vascular cell adhesion molecule 1 expression on cells of the palisading layer in the rheumatoid nodule. CONCLUSION: The presence of similar proinflammatory cytokines and cell adhesion molecules in the rheumatoid nodule and synovial membrane suggests that similar pathogenic processes result in the chronic inflammation and tissue destruction in these lesions.     

Flagellin, a major protein present in SDS-PAGE profiles of Sarkosyl-OMP-enriched fractions from Bordetella bronchiseptica Bvg- or modulated Bvg+ strains

The bvg or vir locus positively regulates the expression of many Bordetella virulence-associated determinants (encoded by vag genes), including cell envelope proteins, in response to environmental stimuli. On the other hand, several genes named vrg genes are negatively controlled by the bvg regulon (Knapp and Mekalanos, 1988). Flagellin is encoded by a vrg gene, which is expressed when the principal virulence factors are eliminated during antigenic modulation or in phase variants (Akerley et al., 1992). We have previously analyzed SDS-PAGE profiles of Sarkosyl-outer membrane protein (OMP)-enriched fractions from B. bronchiseptica Bvg- and modulated Bvg+ strains and reported a major band associated with the avirulent phenotype (Passerini de Rossi et al., 1995). In order to characterize this band we have purified flagellar filaments from Bvg- and modulated Bvg+ strains, and analyzed them by SDS-PAGE. These profiles revealed a single major band of 40 or 45 kDa depending on the strain. The N-terminal amino acid sequence of the putative flagellin expressed by BB7200a was identical over the first 21 residues analyzed to that of the flagellin from the modulated strain BB7865 reported by Akerley et al. (1992). Comparison of the SDS-PAGE profile of flagellar filaments with that of the OMP-enriched fraction of the corresponding strain showed that the flagellum-associated polypeptide had the same electrophoretic mobility as that of the characteristic band of the avirulent phenotype. Furthermore, this band was absent in the OMP-enriched fraction profile from a Bvg- strain subjected to a treatment that removes flagella. Our results indicate that the major protein observed in SDS-PAGE profiles of Sarkosyl-OMP-enriched fractions from B. bronchiseptica Bvg- and modulated Bvg+ strains corresponds to flagellin.     

Mental health pathways from interpersonal violence to health-related outcomes in HIV-positive sexual minority men.

Objective: We examined mental health pathways between interpersonal violence (IPV) and health-related outcomes in HIV-positive sexual minority men engaged with medical care. Method: HIV-positive gay and bisexual men (N = 178) were recruited for this cross-sectional study from 2 public HIV primary care clinics that treated outpatients in an urban setting. Participants (M age = 44.1 years, 36% non-White) filled out a computer-assisted survey and had health-related data extracted from their electronic medical records. We used structural equation modeling to test associations among the latent factors of adult abuse and partner violence (each comprising indicators of physical, sexual, and psychological abuse) and the measured variables: viral load, health-related quality of life (HRQOL), HIV medication adherence, and emergency room (ER) visits. Mediation was tested for the latent construct mental health problems, comprising depression, anxiety, symptomatology of posttraumatic stress disorder, and suicidal ideation. Results: The final model demonstrated acceptable fit, χ2(123) = 157.05, p = .02, CFI = .95, TLI = .94, RMSEA = .04, SRMR = .06, accounting for significant portions of the variance in viral load (13%), HRQOL (41%), adherence (7%), and ER visits (9%), as well as the latent variable mental health problems (24%). Only 1 direct link emerged: a positive association between adult abuse and ER visits. Conclusions: Findings indicate a significant role of IPV and mental health problems in the health of people living with HIV/AIDS. HIV care providers should assess for IPV history and mental health problems in all patients and refer for evidence-based psychosocial treatments that include a focus on health behaviors. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Predictors and pathways from infancy to symptoms of anxiety and depression in early adolescence.

Data from a prospective 11-year longitudinal survey were used to identify early predictors and pathways to symptoms of anxiety and depression at 12–13 years of age, and to examine whether there were unique predictors of anxious versus depressive symptoms. Structural equation modeling was used to explore longitudinal relations between contextual (maternal distress, family adversities, and social support) and temperamental (shyness and emotionality) risk factors in their prediction of informant-consistent symptoms of anxiety and depression. The results show that early risk factors can explain 38% of the variance in boys’ covarying symptoms of anxiety and depression in early adolescence, and 25% of variance in girls’ covarying symptoms. Two main pathways were identified. One pathway was through temperament, as nearly all risk factors were partly mediated through child emotionality in midchildhood. Another pathway was through early contextual risk factors, with all direct and indirect contextual impact from before 5 years of age. Family adversity uniquely predicted depressive symptoms. These findings underscore the persisting impact of contextual predictors in families with children less than 5 years of age. The importance of early interventions to prevent adolescent internalizing problems is stressed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Different cytokine profile and antigen-specificity repertoire in Helicobacter pylori-specific T cell clones from the antrum of chronic gastritis patients with or without peptic ulcer

Helicobacter pylori (Hp) infection almost invariably results in chronic antral gastritis, but only a proportion of patients develop peptic ulcer. Some Hp strains may be more ulcerogenic than others, but some ulcerogenic mechanisms may also depend on the type of the host immune response. In this study, the antigen specificity and the cytokine profile of 53 Hp-specific CD4+ T cell clones derived from the antral mucosa of five patients with Hp-induced uncomplicated chronic gastritis (CG) were assessed and compared with those of 34 Hp-specific CD4+ T cell clones derived from six Hp-infected patients with chronic gastritis and peptic ulcer (CG-PU). The majority (28/34; 82%) of gastric Hp-specific T cell clones from CG-PU patients expressed the Th1 profile and 17 (all Th1) of the 34 clones were specific for cytotoxin-associated protein (CagA). In contrast, 34 (64%) of the 53 Hp-specific gastric T cell clones derived from CG patients were able to secrete both Th1 and Th2 cytokines (Th0 profile) and only 36% expressed a polarized Th1 profile. The majority (85%) of Hp-specific clones from CG patients recognized Hp antigens other than CagA, since 13/53 (25%) were specific for urease, 6 (11%) for VacA, 6 (11%) for HSP and 20 (38%) for other undefined Hp antigens. Results provide evidence that the type of T helper cell response against Hp may vary according to the antigen involved and suggest that a polarized Th1 response may play a role in the genesis of peptic ulcer, whereas a local Th0 response, including interleukin-4 production, may represent an individual host factor which contributes to lower the degree of gastric inflammation and prevent ulcer complication.     

WISC-IV profiles in children with traumatic brain injury: Similarities to and differences from the WISC-III.

The Wechsler Intelligence Scale for Children—Fourth Edition (WISC–IV; D. Wechsler, 2003a) is often utilized to assess children with traumatic brain injury (TBI), although little information is available regarding its psychometric properties in these children. The current study examined WISC–IV performance in a sample of 61 children with TBI. As compared to the standardization sample, results indicated that the TBI group exhibited relative deficits on all subtest and index scores, with the greatest deficits on the Processing Speed Index (PSI) and Coding subtest scores. However, the Perceptual Reasoning Index score was not uniquely sensitive to brain injury, and the Cognitive Processing Index score was less sensitive to TBI than the PSI score. Also, the PSI did not uniquely predict learning and memory abilities, as had been reported in previous studies of the Wechsler Intelligence Scale for Children—Third Edition (WISC–III; D. Wechsler, 1991). The present findings indicate substantive differences between the WISC–III and WISC–IV profiles of children with TBI. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

A family of bicistronic vectors to enhance both local and systemic antitumor effects of HSVtk or cytokine expression in a murine melanoma model

The herpes simplex virus-thymidine kinase/ganciclovir (HSVtk/GCV) system produces both direct and immune-mediated tumor cell killing. Here, we compare the efficacy of HSVtk/GCV with cytokines, alone and in combination, on the tumorigenicity and immunogenicity of B16 cells. With respect to single gene modifications, only HSVtk/GCV, or high-level interleukin-2 (IL-2) secretion, completely prevented tumor growth, whereas granulocyte-macrophage colony-stimulating factor (GM-CSF) generated the best levels of long-term systemic protection. To augment both local killing and immune activation, we constructed bicistronic constructs that express HSVtk and a cytokine within the same cell. Co-expression of HSVtk with IL-2 or GM-CSF enhanced the local antitumor activity of any gene alone. In a tumor-prevention model, HSVtk killing, in an environment preprimed with GM-CSF, generated the best long-term immune protection. However, in a short-term therapy model, continued IL-2 expression was most effective against 3-day established tumors. This probably reflects differences in the activities of IL-2 and GM-CSF in generating short-term, nonspecific immune stimulation compared to long-term immunological memory, respectively. As a prelude to in vivo delivery experiments, we also demonstrated that these bicistronic cassettes can be packaged normally into retroviral (5 x 10(5) virus/ml from pooled populations) and adenoviral vectors (5 x 10(9) virus/ml) and function as predicted within virally infected cells. This family of bicistronic vectors can be used to stimulate synergy between suicide and cytokine genes, overcomes the problems of delivering two genes on separate vectors, and should allow easier preparation of vectors for the delivery of multiple genes to patients' tumor cells.     

CD14-dependent and CD14-independent signaling pathways in murine macrophages from normal and CD14 knockout mice stimulated with lipopolysaccharide or taxol

The antitumor agent, Taxol, shares with bacterial LPS the ability to activate murine macrophages, and its LPS-mimetic effects are blocked by LPS analogue antagonists. Since CD14 is central to the recognition of LPS by macrophages, we sought to examine a role for CD14 in the response to Taxol vs LPS. A comparison of responses of macrophages from wild-type mice with those from mice lacking CD14 due to a targeted disruption of the CD14 gene (CD14-deficient knockout (CD14KO)) revealed that like LPS, Taxol induces both CD14-dependent and -independent pathways of gene activation, although the CD14 dependency of Taxol stimulation is much less striking than that observed with LPS. The macrophage interaction with low concentrations of LPS (< or = 10 ng/ml) is largely CD14 dependent, as evidenced by the lack of induction of TNF-alpha, IL-1beta, and interferon-inducible protein-10 (IP-10) genes by CD14KO macrophages cultured in the absence of soluble CD14 (i.e., in autologous CD14KO -/- mouse serum). However, at high concentrations of LPS or Taxol, a CD14-independent pathway of activation is observed: this pathway leads to minimal IP-10 gene induction, even though induction of TNF-alpha and IL-1beta occurs. Measurements of TNF secretion followed a similar pattern to that observed at the level of steady state mRNA. These data suggest the existence of two pathways of activation by both LPS and Taxol: one that is CD14 dependent and leads to induction of TNF-alpha, IL-1beta, and IP-10 gene induction, and a CD14-independent pathway that results in the induction of TNF-alpha and IL-1beta, with minimal induction of IP-10.     

Descending pathways from the medial longitudinal fasciculus and lateral vestibular nucleus to tail motoneurons in the decerebrate cat

A group of 120 male workers, employed in copperworks (mean age = 41.5 years; mean exposure duration = 17,9 years) at workposts with the highest level of exposure to lead, were covered by the study. Blood levels of the following heavy metals were measured in all workers: Pb, Cd, Mn, Cu, Zn, Ca, Mg as well as concentrations of FEP and GSH, SOD activity in erythrocytes, parameters of lipid metabolism: total cholesterol, HDL2- HDL3-cholesterol, triglycerides, lipid peroxides (LPO), and lecithin-cholesterol acyltransferase (LCAT) activity. Mean blood lead level accounted for 251,86 micrograms/l, and mean level of FEP was slightly above normal. That may indicate moderate lead deposits in smelters. Concentrations of other metals remained within normal limits. No significant disturbances in lipid metabolism were observed. Along with expected positive correlation between lead blood level and FEP, a significant negative correlation between lead and cholesterol levels as well as between FEP and serum cholesterol was found. Moreover, a significant negative correlation between FEP and serum LPO, as well as a significant positive correlation between concentration and HDL2-cholesterol level and between FEP concentration and SOD activity in erythrocytes were noted. We believe that unexpected outcome of our investigations could result from the adaptation of healthy smelters to the environmental conditions. It is assumed that further exposure could weak antioxidant mechanisms and lead, in consequence, to the manifestation of symptoms induced by harmful effect of free radicals.     

Change in susceptibility of group B streptococci to penicillin G from 1968 through 1975

This report describes a study of 212 isolates of group B streptococci from sore throats over an 8-year period. A small but increasing percentage showed increased resistance to penicillin G when tested in an in vitro system.     

Failure to control prepotent pathways in early stage dementia of the Alzheimer's type: Evidence from dichotic listening.

The authors examined the right ear advantage in a dichotic listening task in healthy aging and very mild and mild stages of Alzheimer's disease. Subjects were simultaneously presented 3 pairs of digits to the left and right ears (e.g., left ear: 4, 3, 1; right ear: 9, 2, 5) for immediate ordered recall. Four lists of triads were presented, varying in presentation rate between digit pairs within a triad (0.5, 1.0, 1.5, 2.0 s). Results indicated that the very mild and mild Alzheimer's groups showed a larger right ear advantage in free recall compared with the healthy controls, indicating a tendency to respond to the prepotent left hemisphere pathway for language processing. Also, the right ear advantage and proportion of switches made during recall were correlated with psychometric measures of frontal lobe function in the mild Alzheimer's group but not in the very mild or healthy control groups. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Polysynaptic neuronal pathways from group I and group II afferents innervating tail muscles to hindlimb motoneurons in the cat

Postsynaptic potentials evoked in motoneurons innervating m. posterior biceps and semitendinosus (PBSt) and m. triceps surae (GS) by low threshold afferents from various tail muscles located at the level of the second-third caudal vertebrae were investigated in the non-anesthetized and spinalized cat. Afferent inputs from tail muscles on both sides predominantly evoked depolarizing potential in PBSt motoneurons and hyperpolarizing potential in GS motoneurons. The findings suggest that in general, tail muscle afferents facilitate flexor and inhibit extensor hindlimb motoneurons through polysynaptic pathways, so that the pelvic girdle is kept in a low position to maintain the stability of the body irrespective of different movements or posture of the tail.     

Gender control and Olympics events from Albertville to Atlanta or good practices in gene biochemistry

A tendon forms an integral part of musculotendinous unit. It transmits the tensions generated in muscles to bone. Tendons are stronger than muscles and are able to withstand larger forces. Tendons are subject to both tensile and high compressive forces.     

Control of heart rate during transition from intravenous to oral diltiazem in atrial fibrillation or flutter

We tested whether patients presenting with atrial fibrillation (AF) or flutter (AFl) with a rapid ventricular response could maintain control of heart rate while transferring from a bolus and continuous infusion of intravenous diltiazem to oral diltiazem. Forty patients with AF or AFI and sustained ventricular rate > or = 120 beats/min received intravenous diltiazem "bolus" (20 to 25 mg for 2 minutes) and "infusion" (5 to 15 mg/hour for 6 to 20 hours). Oral long-acting diltiazem (diltiazem CD 180, 300, or 360 mg/24 hours) was administered in patients in whom stable heart rate control was attained during constant infusion. Intravenous diltiazem infusion was discontinued 4 hours after the first oral dose, and patients were monitored during 48 subsequent hours of "transition" to oral therapy. Response to diltiazem was defined as heart rate <100 beats/min, > or = 20% decrease in heart rate from baseline, or conversion to sinus rhythm. Other rate control or antiarrhythmic medications were not allowed during the study period. Thirty-seven of 40 patients maintained heart rate control during the bolus, and 35 of the remaining 37 maintained control during the infusion of intravenous diltiazem. Of the 35 patients achieving heart rate control with intravenous diltiazem who entered the transition to oral therapy, 27 maintained heart rate control (response rate of 77%/, 95% confidence interval 63% to 91%). The median infusion rate of intravenous diltiazem was 10 mg/hour, and the median dose of oral diltiazem CD was 300 mg/day. Oral long-acting diltiazem was 77% effective in controlling ventricular response over 48 hours in patients with AF or AFl in whom ventricular response was initially controlled with intravenous diltiazem.