Association Between Metacognition and Mood Symptoms Poststroke.

INTRODUCTION: The link between metacognition and mood has been well established, particularly in other conditions with psychological comorbidity, however, there is no evidence regarding this association in the area of stroke. AIM: The aim of this study was to examine the association between metacognition, based on the Self-Regulatory Executive Function model, and mood symptoms in the acute phase after stroke. METHODS: One hundred thirty patients were recruited to a prospective stroke study in Bahrain, and n = 64 were assessed for mood and cognition. A neuropsychological battery of cognitive assessments included the following measures: the Mini-Mental State Examination, the Trail Making Test (A+B), and the Metacognition Questionnaire 30 (MCQ-30) for metacognition. The Hospital Anxiety and Depression Scale assessed mood symptoms, and stroke severity was measured using the National Institute of Health Stroke Severity Scale. RESULTS: Total MCQ-30 scores were significantly associated with both anxiety (r = .47, P = .001) and depression (r = .54, P <. 0001). The MCQ-30 subscales' cognitive confidence, cognitive self-consciousness, and uncontrollability/danger were the specific factors to be associated with mood symptoms (P < .01). Global cognition (r =.32, P < .01), but not executive function, was significantly associated with depression only. Metacognition remained a statistically significant correlate with depression (ß = .42, P < .0001) and anxiety (ß = .51, P < .0001) after adjusting for education and global cognition. DISCUSSION: Metacognition is a better determinant of mood symptoms after stroke, especially in regions where illiteracy levels are high in older populations, in comparison to executive function and global cognition.

Selective newborn screening of inborn errors of amino acids, organic acids and fatty acids metabolism in the Kingdom of Bahrain.

Mandatory newborn screening for metabolic disorders has not been implemented in most Middle Eastern countries. Early detection and treatment of inborn errors of metabolism can reduce mortality and minimize morbidity. Preliminary studies conducted in some parts of Middle East suggest that the incidences of inborn errors of metabolism are reported to be higher in the region than anywhere else in the world due to the consanguinity. In this study the incidence of inborn errors of amino acids, organic acids and fatty acids oxidation disorders was investigated from the results of blood spot analysis of 1986 symptomatic children from 1st January 2008 to 31st of December 2011. Out of 1986 newborns screened 25 infants were diagnosed and confirmed with amino acids (n=11), organic acids (n=9) and fatty acids oxidation (n=5) disorders. Overall incidences based on number of live birth between 2008 and 2011 inclusive were 1:6000, 1:8000 and 1:14,000 for amino acids, organic acids and fatty acids oxidation disorders; respectively. Out of 25 infants diagnosed, 21 were the children of first cousin marriages. Results from this study suggest high incidence of inborn errors of amino acids, organic acids and fatty acids oxidation metabolism in Bahrain and significant contribution of consanguinity in inherited metabolic disorders. Mandatory screening for inborn errors of metabolism in Bahrain is highly recommended.

RANTES promotes growth and survival of human first-trimester forebrain astrocytes.

We have examined the role of alpha and beta chemokines in the promotion of the ontogenetic development of the brain. RANTES was expressed preferentially in human fetal astrocytes in an age-dependent manner. Astrocytes from 5-week-old brains showed high proliferation and reduced survival, whereas 10-week-old astrocytes exhibited opposite effects. These effects were suppressed by anti-RANTES or anti-RANTES receptor antibodies and were enhanced by recombinant RANTES. RANTES induced tyrosine phosphorylation of several cellular proteins and nuclear translocation of STAT-1 in astrocytes. Interferon-gamma (IFN-gamma) was required for RANTES effects because RANTES induced IFN-gamma and only 10-week-old astrocytes expressed the IFN-gamma receptor. Blocking of IFN-gamma with antibody reversed the effects of RANTES, indicating that cytokine/chemokine networks are critically involved in brain development.

The gene for a T lymphocyte triggering factor from African trypanosomes.

An early and essential event in the protective immune response against most viruses and protozoa is the production of interferon-gamma (IFN-gamma). In contrast, during infection with African trypanosomes, protozoan parasites that cause human sleeping sickness, the increased levels of IFN-gamma do not correlate with a protective response. We showed previously that African trypanosomes express a protein called T lymphocyte triggering factor (TLTF), which triggers CD8(+) T lymphocytes to proliferate and to secrete IFN-gamma. Here, we isolate the gene for TLTF and demonstrate that the recombinant version of TLTF specifically induces CD8(+), but not CD4(+), T cells to secrete IFN-gamma. Studies with TLTF fused to the green fluorescent protein show that TLTF is localized to small vesicles that are found primarily at or near the flagellar pocket, the site of secretion in trypanosomes. TLTF is likely to be only the first example of a class of proteins that we designate as trypanokines, i.e., factors secreted by trypanosomes that modulate the cytokine network of the host immune system for the benefit of the parasite.

Both CD4+ and CD8+ T cells are essential to induce experimental autoimmune myasthenia gravis.

CD4+ T cells have been shown to be crucial in the development of experimental autoimmune myasthenia gravis (EAMG). The role of CD8+ T cells in EAMG is less well established. We previously showed that antibody depletion of CD8+ T cells in rats effectively suppresses EAMG. To further study the role and relationship of CD4+ versus CD8+ T cells in induction of EAMG, CD4-/-, CD8-/-, and CD4-8- mutant C57BL/6 mice and the parent CD4+8- wild-type mice were immunized with Torpedo acetylcholine receptor (AChR) plus complete Freund's adjuvant. Clinical EAMG was nearly completely prevented in CD4-8-, CD4-/-, and CD8-/- mice. This was associated with strongly reduced AChR-specific T and B cell responses, and with reduced levels of AChR-reactive interferon gamma (IFN-gamma) and interleukin 4 (IL-4) mRNA-expressing cells in lymphoid organs when compared with CD4+8+ wild-type mice. We conclude that (a) both CD4+ and CD8+ T cells are essential for development of EAMG, and a collaboration between these cell types may be necessary; (b) CD4+ as well as CD8+ T cells secrete IFN-gamma and IL-4, and both cytokines are involved in the development of EAMG; and (c), besides T cells, other immune cells might also be responsible for help of anti-AChR antibody production.

Brucellosis in camels, cattle and humans: associations and evaluation of serological tests used for diagnosis of the disease in certain nomadic localities in Sudan.

Brucellosis was studied in 2,225 camels, 20 camel nomads and 33 abattoir workers in certain nomadic localities in Sudan, using serum and milk samples. Lymph nodes, testicular tissues and udder tissues from positive camels and hygroma aspirates from three affected cows were used for isolation of Brucella. Serum samples were examined by Rose Bengal plate test (RBPT), modified RBPT (mRBPT), serum agglutination test (SAT) and competitive enzyme-linked immunosorbent assay (cELISA), and milk by the milk ring test. Overall seroprevalence in camels (milk and serum samples) was 37.5%. The seroprevalence in males was 28.2% and in females 40.1%. Twelve (60%) of the 20 nomads and three (9%) of the 33 abattoir workers had positive antibody titres. Brucella abortus biovar 6 was isolated from two camels and three cows. Two isolates, one from each species, were atypical. The bacteriological findings suggested that camels were infected from cattle, the primary hosts of B. abortus. The mRBPT was suitable for screening camel sera for brucellosis, but the cELISA detected 2.1% more positives. The SAT antibody concentrations ranged between < 13 and 3,282 IU/ml.

Associations between ApoE gene and psychological consequences post stroke in a Bahraini cohort.

BACKGROUND: The contribution of genetic factors such as the presence of ApoE allele e4 and its association with psychological consequences post stroke remains unknown within Middle-Eastern regions. This study examined the association of ApoE genotype with cognitive impairment and mood in stroke patients and compare with healthy older adults in Bahrain. METHOD: A prospective sample of n = 62 stroke patients (case group) and n = 53 healthy ageing individuals (control group) were eligible to participate in the study. A neuropsychological battery of cognitive assessments were conducted on all participants, and then stratified by cognitive function: no cognitive impairment, mild cognitive impairment and moderate to severe cognitive impairment. Anxiety and depression were assessed using the Hospital Anxiety and Depression Scale (HADS). RESULTS: Most frequent ApoE genotype was e2/e3 in case (44%) and control groups (63%). ApoE allele e3 had the highest frequency for both groups with all stroke patients presenting with this allele and 86% for the control group (χ2 = 12.14, p < .0001). Stroke patients' non-carriers for ApoE allele e4 performed better on all cognitive measures but differences were not statistically significant (ns). Carriers of ApoE allele e2 in both groups had less mood symptoms compared to non-carriers. DISCUSSION: ApoE genotype e3/e4 and e4/e4 was low in this Bahraini cohort explaining why there may been no significant associations found for this genotype variant with cognitive impairment. Further investigation of cognitive impairment and mood dysregulation with the different variants of the ApoE gene in general ageing and stroke populations is required from different ethno-cultural groups and geographical regions globally.

Mutations in the promoter of adenylyl cyclase (AC)-III gene, overexpression of AC-III mRNA, and enhanced cAMP generation in islets from the spontaneously diabetic GK rat model of type 2 diabetes.

Glucose-induced insulin release is decreased in the spontaneously diabetic GK rat, a nonobese rodent model of type 2 diabetes. Forskolin restores the impaired insulin release in both the isolated perfused pancreas and isolated islets from these rats (Abdel-Halim et al., Diabetes 45:934-940, 1996). We demonstrate here that the insulinotropic effect of forskolin in the GK rat is due to increased generation of cAMP and that it is associated with overexpression of adenylyl cyclase (AC)-III mRNA and gene mutations. The AC-III mRNA overexpression was demonstrated by in situ hybridization using oligonucleotide probes binding to different regions of the rat AC-III mRNA. It was associated with the presence of two point mutations identified at positions -28 bp (A --> G) and -358 bp (A --> C) of the promoter region of the AC-III gene and was demonstrable in both GK rat islets and peripheral blood cells. Transfection of COS cells with a luciferase reporter gene system revealed up to 25-fold increased promoter activity of GK AC-III promoter when compared with normal rat promoter (P < 0.0001). In conclusion, forskolin restores the impaired insulin release in islets of the GK rat through enhanced cAMP generation. This is linked to overexpression of AC-III mRNA in GK islets due to two functional point mutations in the promoter region of the AC-III gene.

Lovastatin inhibits interferon-gamma-induced Trypanosoma brucei brucei proliferation: evidence for mevalonate pathway involvement.

Interferon-gamma (IFN-gamma) is an essential immunoregulating molecule that has recently been shown to have a growth stimulatory effect on Trypanosoma brucei brucei (T. b. brucei). The signalling pathway(s) involved during this triggering are unknown. Since the different products from the biosynthesis pathway utilizing mevalonate have several important cellular functions, ranging from cholesterol synthesis to growth control, we here investigate the possible role for the mevalonate pathway in IFN-gamma-driven parasite proliferation. Thus, lovastatin, a hydroxymethylglutaryl-coenzyme A (HMG-CoA) reductase-inhibiting drug, was incubated at different concentrations in vitro with T. b. brucei. The parasites were then stimulated with a broad concentration range of rIFN-gamma. The effect on proliferation or growth was measured either by the tritium-labeled thymidine incorporation assay or by direct counting of parasites from the cultures using light microscopy. The maximum proliferative response was obtained with IFN-gamma at a concentration of 10(3) U/ml added to 10(6) parasites. This response was markedly decreased with lovastatin, even at a low concentration (0.1 mM). The effect of lovastatin was reversed by the addition of 10 mM mevalonate. IFN-gamma at a concentration of 10(4) U/ml showed no proliferative effect. Addition of mevalonate to this concentration of IFN-gamma gave a threefold increase in parasite proliferation. Our data suggest that a low concentration of IFN-gamma induces parasite growth, a high concentration has the opposite effect, and both these events are regulated by activity or inactivity of the mevalonate pathway.

Orthodontic movement induces high numbers of cells expressing IFN-gamma at mRNA and protein levels.

Cytokines are important signaling proteins that are liberated during immune challenges and exhibit many modulatory activities. However, their role in periodontal modeling during orthodontic tooth movement is not fully understood. The aim of this study was to analyze effects of mechanical force during orthodontic tooth movement, in the pressure zone, on the induction of interferon-gamma (IFN-gamma) as a proinflammatory cytokine of Th1 type and interleukin-4 (IL-4)/IL-10 as anti-inflammatory cytokines of Th2 type. In 12 Wistar rats 40-45 days old, the maxillary first molar was moved mesially by means of a closed coil spring for 3, 7, and 10 days. The contralateral side served as a control. IFN-gamma, IL-4, and IL-10 mRNA were determined by in situ, hybridization, and protein levels of IFN-gamma was measured by immunohistochemistry. Induction of IFN-gamma at both mRNA and protein levels was significantly higher on the experimental side than on the contralateral control side on day 3. The signal gradually became stronger on day 7 and remained high on day 10. Cytokines of the Th2 type (IL-4 and IL-10) were not detected at all examined time points in both pressure and contralateral control sides. Considering the potential immunoregulatory roles played by IFN-gamma, our data suggest that IFN-gamma may be involved in periodontium remodeling during orthodontic tooth movement.

Chemokines are upregulated during orthodontic tooth movement.

In the early stage of orthodontic tooth movement, an acute inflammatory response characterized by the migration of leukocytes occurs. This response suggests the presence of specific chemotactic signals that may play a role in the mechanism of bone remodeling, in particular in resorption. The aim of the present study was to explore the induction of potential chemokines at the resorption side during orthodontic tooth movement. Monocyte chemoattractant protein-1 (MCP-1), regulated on activation normal T cell expressed and secreted (RANTES), and macrophage inflammatory protein-2 (MIP-2) were examined by in situ hybridization using radioactive synthetic oligoneucleotide probes. Mesial movement of the upper first molars was performed with a fixed appliance for 3, 7, and 10 days. The results demonstrated that MCP-1, RANTES, and MIP-2 were highly expressed during orthodontic movement. On day 3, MCP-1 showed maximum induction in the pressure zone, followed in intensity by RANTES and MIP-2, although not in the contralateral control side. The induction of these chemokines had declined on day 7 and reached low levels on day 10. Our data suggest that chemokines are induced early in the application of force, and such induction may contribute to the early inflammatory response that may be responsible in part for the ensuing bone remodeling.

A novel mechanism for cytokine regulation: screening, selection, and characterization of anticytokine monoclonal and polyclonal autoantibodies.

We have recently described an immunoregulatory mechanism involving release of neutralizing autoantibodies (Aab) to cytokines during bacterial infections. Intraperitoneal inoculation of Haemophilus influenzae type b (Hib) into Sprague-Dawley rats resulted in high levels of inflammatory mediators early after infection. Increased titers of cytokine Aab were observed, with a peak at day 7. We cloned Aab-producing B cells. Screening of the clones with five different cytokines resulted in detection of Aab-producing clones reactive with each cytokine. After repeated subcloning, monoclonal Aab (mAab) were selected and characterized for their specificity, isotypes, and affinities. To elucidate regulatory importance, mAab to interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) dose-dependently inhibited IFN-gamma-induced MHC expression by peritoneal macrophages and TNF-alpha-induced thymocyte proliferation, respectively. Fab fragments exhibited binding and neutralizing effects, confirming specificities. Cross-reactivity with other rat cytokines was excluded. Pools of clones containing several mAab to each cytokine were obtained and served as polyclonal Aab. The relative affinity of the Aab was determined and found to be of high index. The characterized Aab were tested in methodologic assays for cytokine detection, revealing that some Aab were useful in a cell release capturing (CRC) ELISA.

Induction of interferon-gamma, transforming growth factor-beta, and interleukin-4 in mouse strains with different susceptibilities to Trypanosoma brucei brucei.

A Trypanosoma brucei brucei-derived lymphocyte triggering factor (TLTF) induced CD8+ T cells to produce IFN-gamma, which in turn stimulates parasite growth. This parasite-host interaction was studied in mouse strains that are either relatively susceptible (C3H/He) or resistant (C57Bl/6J) to infection, as well as in athymic nude mice. In all mouse strains, T. b. brucei infection caused a strong induction of IFN-gamma production by spleen mononuclear cells (MNC). In vivo blocking of IFN-gamma by intraperitoneal injection of mouse monoclonal anti-IFN-gamma antibody suppressed parasite growth and increased survival of both C3H/H3 and C57Bl/6J animals, suggesting that, irrespective of strain-related disease susceptibility, IFN-gamma is a growth-promoting stimulus for T. b. brucei. Spleen MNC from noninfected mice of all strains were in vitro like-wise strongly induced to IFN-gamma production when exposed to TLTF. This suggests that CD8+ expressing T cell receptor (TCR) alpha/beta, gamma/delta-bearing T cells and NK cells may all be triggered to IFN-gamma production by TLTF. In all mouse strains, TLTF also caused an increase in the number of cells expressing mRNA for TGF-beta in vitro. However, significant triggering to IL-4 mRNA expression only occurred in the relatively disease-resistant C57Bl/6J strain. As IL-4 is required for the synthesis and class switches of immunoglobulins, which are essential host immune defenses against T. b. brucei, the degree of resistance may be related to inherent strain ability to produce IL-4 in response to TLTF.

Chemokines are produced in the brain early during the course of experimental African trypanosomiasis.

African trypanosomiasis is characterized by progressive central nervous system (CNS) involvement. Using single and double immunohistochemistry, we evaluated the induction of alpha- and beta-chemokines in brains of Sprague-Dawley rats infected with Trypanosoma brucei brucei (T. b. brucei) and identified their cellular source. The results showed high production of MIP-2, RANTES and MIP-1alpha and to a lower extend MCP-1 in infected animals compared to controls. MIP-2, RANTES and MIP-1alpha were produced early by astrocytes and microglia and later by macrophages and T-cells. These findings suggest that chemokines may contribute to the immunopathogenesis that occurs in the CNS early during infections.

Gamma interferon expression and major histocompatibility complex induction during measles and vesicular stomatitis virus infections of the brain.

Lymphocytic interferon gamma (IFN-gamma) production and major histocompatibility complex (MHC) antigen induction were studied in experimental measles and vesicular stomatitis virus infections in the brain. Fifteen-day-old Sprague-Dawley rats injected intracerebrally with the HNT strain of measles virus showed already within 1 day after infection an increased number of cells producing IFN-gamma in the spleen, cervical lymph nodes and leptomeninges. These rats recovered after a transient neuronal infection in the brain. Rats infected intracerebrally with vesicular stomatitis virus, on the other hand, all succumbed after 2 days and showed no IFN-gamma production in lymphoid cells. Immunohistochemically MHC class I antigen appeared in infected and uninfected cells in the brain during replication of both viruses. A role for the recently discovered nerve fibres with IFN-gamma-like immunoreactivity, which are normally present in the brain, in the MHC antigen induction is discussed.

Different trypanozoan species possess CD8 dependent lymphocyte triggering factor-like activity.

Trypanosoma brucei brucei (T. b. brucei) release a molecule, trypanosome derived lymphocyte triggering factor (TLTF), which stimulates CD8+ cells to produce cytokines and to proliferate. We now report that T. evansi, T.b. gambiense and T.b. rhodesiense also contain factor(s) with similar activity. Thus, homogenates from these parasite taxa triggered mouse or rat lymphoid tissue of mononuclear cells (MNC) to produce interferon gamma (IFN-gamma) and to proliferate. These responses were dependent on CD8 since the activity was blocked by (a) anti CD8 antibodies, (b) occurred in CD8+ but not in CD8- mice and (c) was recorded in human CD8+ but not in CD4+ enriched peripheral blood mononuclear cells (PBL). The presence of TLTF or TLTF-like molecules in the trypanozoan species was also examined by T.b. brucei directed anti-TLTF Mabs using two Mabs with inhibitory activity and one with stimulatory activity. The lymphocyte triggering activity of T.b. gambiense and T.b. rhodesiense, but not T. evansi, was affected by the anti-TLTF Mabs. We conclude that T. evansi, T.b. rhodesiense and T.b. gambiense similar to T.b. brucei, all possess molecule(s) which CD8 dependently trigger lymphocytes. The latter three, related parasite taxa, share TLTF antibody binding epitopes.

Tyrosine kinases are required for IFN-gamma-induced growth of human embryonic forebrain astrocytes.

The cytokine IFN-gamma was shown to regulate growth and differentiation of human embryonic forebrain astrocytes. This work investigated a signalling pathway used by IFN-gamma during the process of growth regulation of human fetal astrocytes obtained from first trimester embryos. IFN-gamma induced significantly higher cell survival compared to that of unexposed cultures, and this survival could be suppressed by incubation with the tyrosine protein kinase (TPK) specific inhibitor tyrphostin A47 at the non-toxic concentration of 10(-6) M. The signal transducer and activator of transcription (STAT)-1 was translocated into the nucleus upon IFN-gamma stimulation, which was also blocked by incubation with A47. Our data demonstrate that TPKs are actively involved in growth regulation of the developing brain astrocytes induced by IFN-gamma.

Expression of interleukin-6 in human dorsal root ganglion cells.

The expression of Interleukin-6 (IL-6) was studied in normal dorsal root ganglia (DRG) of juvenile and foetal humans, using immunohistochemistry and in situ hybridization techniques. There was an expression of IL-6-like immunoreactivity in more than 75% out of neuronal cells in the juvenile ganglia with a peripheral localization, and also an expression in the foetal ganglion cells. There was a co-localization of IL-6 with substance P (SP) and calcitonin gene-related peptide (CGRP) in more than 60% of the DRG cells, respectively. By in situ hybridization 0.9% of the cells in the juvenile ganglia and 1.1% of the cells in the foetal ganglia showed a positive signal for IL-6. In addition, expression of IL-6 was found in juvenile medulla spinalis, preferentially in the white matter.

Interactions between human embryonic forebrain cells and the cytokines interferon-gamma and interleukin-4.

In the present study we investigated the capacity of human first trimester forebrain cells at different gestational ages to produce IFN-gamma and IL-4. We also studied the effects of IFN-gamma on their proliferation, survival and expression of MHC antigens. IFN-gamma but not IL-4 was spontaneously produced after 24 h cultures. Furthermore, IFN-gamma exhibited anti-proliferative effects and induced MHC expression on these cells. However, the IFN-gamma exposed cultures showed significantly higher cell survival compared to un-exposed cultures. Co-culture with IL-4 blocked the IFN-gamma production and reversed its anti-proliferative effects. These interactions suggest important roles for cytokines in the survival, proliferation and differentiation of human embryonic and fetal forebrain cells.

Cytokines and anti-cytokine autoantibodies during experimental african trypanosomiasis in mice with disrupted interferon-gamma and interferon-gamma receptor genes.

We studied cytokines and anti-cytokine autoantibodies (Aabs) during T.b.brucei infections in IFN-gamma-/-, IFN-gammaR-/- and wild-type mice. Increased serum levels of IFN-gamma, TNF-gamma and IL-4 with decreased Aabs to these cytokines were recorded early during infections in all mice (except IFN-gamma in IFN-gamma-/- mice). Later, these responses were reversed, and surprisingly Aabs reacting to IFN-gamma in the IFN-gamma -/- mice were detected. To examine the possibility that an IFN-ç immunoreactive molecule might be expressed due to infections and upon gene deletion, anti-IFN-gamma antibody was inoculated and resulted in abrogation of such Aabs. The scenario was different for IL-10 and TGF- since IFN-gammaR-/- and wild-type mice showed low cytokines and high Aabs early during infections, but later high cytokines and low Aabs were registered. Interestingly, IFN-gamma-/- mice exhibited reversed levels of both IL-10 and TGF-beta, and also of their Aabs. Fab fragments of purified serum immunoglobulins showed binding and neutralizing effects in biological assays. Pre-absorption of the Fab fragments with a cytokine inhibited the binding and neutralization effects of this cytokine, but not of other cytokines. These results highlight an important role for autoimmunity in cytokine regulation, and that genomic deletion of IFN-gamma modulates cytokines and their Aab responses in experimental African trypanosomiasis.