Monocyte Function in Parkinson's Disease and the Impact of Autologous Serum on Phagocytosis.

Background: Increasing evidence implicates involvement of the innate immune system in the initiation and progression of Parkinson's disease (PD). Monocytes and monocyte-derived cells perform a number of functions, such as phagocytosis, chemotaxis, and cytokine secretion, which may be particularly relevant to PD pathology. The behavior of these cells in early-moderate disease, in conditions more similar to the in-vivo environment has not been fully evaluated. Research Question: Does monocyte function, including phagocytosis, chemotaxis and cytokine secretion, differ in early-moderate PD compared to age and gender-matched controls? Methods: Participants included PD patients (n = 41) with early-moderate stage disease (Hoehn and Yahr ≤2) and age and gender matched controls (n = 41). Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood and monocytes were further separated using CD14 magnetic beads. Functional assays, including bead phagocytosis (in standard medium and autologous serum), Boyden chamber trans-well chemotaxis, and cytokine secretion on lipopolysaccharide stimulation were performed. Monocyte surface markers relating to chemotaxis were measured using immunohistochemistry and flow cytometry. Between-group analysis was performed using paired t-tests. Results: An autologous serum environment significantly increased bead phagocytosis compared to standard medium as expected, in both patients and controls. When in autologous serum, PD monocytes demonstrated enhanced phagocytosis compared to control monocytes (p = 0.029). The level of serum-based phagocytosis was influenced by complement inactivation and the origin of the serum. There were no significant differences between PD and controls in terms of standard medium based monocyte migration or cytokine secretion in this cohort. Conclusions: Autologous serum has a significant influence on monocyte phagocytosis and reveals increased phagocytic capacity in early-moderate PD compared to controls. These conditions may better reflect the function of monocytes in-vivo in PD patients than standard medium based phagocytosis assays. Further studies will be required to replicate these results in larger cohorts, including earlier and later stages of disease, and to understand which serum factors are responsible for this observation and the potential mechanistic relevance to PD pathogenesis.Funding for this work was provided by Addenbrooke’s Charitable Trust, the Rosetrees Trust and the NIHR Cambridge Biomedical Research Centre. RSW was supported by a Fellowship from Addenbrooke’s Charitable Trust. DKV is supported by a Junior Research Fellowship from Homerton College, Cambridge. KMS is supported by a Fellowship from the Wellcome Trust. CHWG is supported by a Clinician Scientist Fellowship from the Medical Research Council. RAB is an NIHR Senior Investigator and is supported by the Wellcome Trust-MRC Cambridge Stem Cell Institute

Peripheral innate immune and bacterial signals relate to clinical heterogeneity in Parkinson's disease.

The innate immune system is implicated in Parkinson's disease (PD), but peripheral in-vivo clinical evidence of the components and driving mechanisms involved and their relationship with clinical heterogeneity and progression to dementia remain poorly explored. We examined changes in peripheral innate immune-related markers in PD cases (n = 41) stratified according to risk of developing early dementia. 'Higher Risk'(HR) (n = 23) and 'Lower Risk' (LR) (n = 18) groups were defined according to neuropsychological predictors and MAPT H1/H2 genotype, and compared to age, gender and genotype-matched controls. Monocyte subsets and expression of key surface markers were measured using flow cytometry. Serum markers including alpha-synuclein, inflammasome-related caspase-1 and bacterial translocation-related endotoxin were measured using quantitative immuno-based assays. Specific markers were further investigated using monocyte assays and validated in plasma samples from a larger incident PD cohort (n = 95). We found that classical monocyte frequency was elevated in PD cases compared to controls, driven predominantly by the HR group, in whom Toll-Like Receptor (TLR)4+ monocytes and monocyte Triggering Receptor Expressed on Myeloid cells-2 (TREM2) expression were also increased. Monocyte Human Leukocyte Antigen (HLA)-DR expression correlated with clinical variables, with lower levels associated with worse cognitive/motor performance. Notably, monocyte changes were accompanied by elevated serum bacterial endotoxin, again predominantly in the HR group. Serum alpha-synuclein and inflammasome-related caspase-1 were decreased in PD cases compared to controls regardless of group, with decreased monocyte alpha-synuclein secretion in HR cases. Further, alpha-synuclein and caspase-1 correlated positively in serum and monocyte lysates, and in plasma from the larger cohort, though no associations were seen with baseline or 36-month longitudinal clinical data. Principal Components Analysis of all monocyte and significant serum markers indicated 3 major components. Component 1 (alpha-synuclein, caspase-1, TLR2+ monocytes) differentiated PD cases and controls in both groups, while Component 2 (endotoxin, monocyte TREM2, alpha-synuclein) did so predominantly in the HR group. Component 3 (classical monocytes, alpha-synuclein) also differentiated cases and controls overall in both groups. These findings demonstrate that systemic innate immune changes are present in PD and are greatest in those at higher risk of rapid progression to dementia. Markers associated with PD per-se (alpha-synuclein, caspase-1), differ from those related to cognitive progression and clinical heterogeneity (endotoxin, TREM2, TLR4, classical monocytes, HLA-DR), with mechanistic and therapeutic implications. Alpha-synuclein and caspase-1 are associated, suggesting inflammasome involvement common to all PD, while bacterial translocation associated changes may contribute towards progression to Parkinson's dementia. Additionally, HLA-DR-associated variations in antigen presentation/clearance may modulate existing clinical disease

Monocyte Function in Parkinson's Disease and the Impact of Autologous Serum on Phagocytosis

Background: Increasing evidence implicates involvement of the innate immune system in the initiation and progression of Parkinson's disease (PD). Monocytes and monocyte-derived cells perform a number of functions, such as phagocytosis, chemotaxis, and cytokine secretion, which may be particularly relevant to PD pathology. The behavior of these cells in early-moderate disease, in conditions more similar to the in-vivo environment has not been fully evaluated.Research Question: Does monocyte function, including phagocytosis, chemotaxis and cytokine secretion, differ in early-moderate PD compared to age and gender-matched controls?Methods: Participants included PD patients (n = 41) with early-moderate stage disease (Hoehn and Yahr ≤2) and age and gender matched controls (n = 41). Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood and monocytes were further separated using CD14 magnetic beads. Functional assays, including bead phagocytosis (in standard medium and autologous serum), Boyden chamber trans-well chemotaxis, and cytokine secretion on lipopolysaccharide stimulation were performed. Monocyte surface markers relating to chemotaxis were measured using immunohistochemistry and flow cytometry. Between-group analysis was performed using paired t-tests.Results: An autologous serum environment significantly increased bead phagocytosis compared to standard medium as expected, in both patients and controls. When in autologous serum, PD monocytes demonstrated enhanced phagocytosis compared to control monocytes (p = 0.029). The level of serum-based phagocytosis was influenced by complement inactivation and the origin of the serum. There were no significant differences between PD and controls in terms of standard medium based monocyte migration or cytokine secretion in this cohort.Conclusions: Autologous serum has a significant influence on monocyte phagocytosis and reveals increased phagocytic capacity in early-moderate PD compared to controls. These conditions may better reflect the function of monocytes in-vivo in PD patients than standard medium based phagocytosis assays. Further studies will be required to replicate these results in larger cohorts, including earlier and later stages of disease, and to understand which serum factors are responsible for this observation and the potential mechanistic relevance to PD pathogenesis

Bioactive and Antimicrobial Properties of Eggplant (<i>Solanum melongena</i> L.) under Microwave Cooking

Fruits and vegetables constitute a considerable amount of antioxidants and among them eggplant is a rich source of polyphenol compounds. This study investigated the bioactive and antimicrobial properties of eggplant under different degree of microwave cooking. The eggplant was cooked for 7 min (light cooked), 10 min (medium cooked), and 15 min (high cooked). The highest total polyphenol content was observed in the light cooked eggplant sample (27.35 mg gallic acid equivalent (GAE)/g dry weight (DW)) followed by high cooked sample (26.10 mg GAE/g DW), while the lowest total polyphenol content (2.79 mg GAE/g DW) was obtained for the uncooked (control) sample. The total polyphenol content of the samples ranged in the following order; light cooked > high cooked > medium cooked > uncooked. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging of eggplant ranged between 16.16% (control) and 47.88% (high cooked). The highest reducing power was exhibited by the light cooked (absorbance 1.708) eggplant sample followed by the high cooked (absorbance 1.597), while the lowest reducing power was shown by uncooked sample (absorbance 0.389). Moreover, antimicrobial studies showed that light cooked eggplant sample demonstrated broad-spectrum inhibition of growth in Gram-positive as well as Gram-negative bacteria and Candida albicans. Slightly lower antimicrobial potential was exhibited by medium cooked eggplant sample while no antibacterial or antifungal activity was recorded for the extract of high cooked eggplant sample. Microwave cooking might be a method to enhance the antioxidant and antimicrobial potential of eggplant

Different drying techniques effect on the bioactive properties of rose petals

This study explored the effect of different drying methods (sun, shade, oven) on the total polyphenol content (TPC) using the Folin–Ciocalteu (FC) procedure, total flavonoid content (TFC) using the AlCl3 colorimetric method, volatile organic compounds using GC/MS and the antioxidant properties of the rose petals using the protocol of DPPH assy. The results demonstrated that the drying methods had a significant impact and the highest TPC (34.24 mg gallic acid equivalent (GAE)/ g fresh weight (FW)) and TFC (5.56 mg catechin equivalent (CE)/g FW) were obtained for the oven-dried sample. While, the fresh sample exhibited the lowest TPC (15.6 mg GAE/g FW) and TFC (3.83 mg CE/g FW), respectively. Similarly, the oven-dried sample showed the highest DPPH scavenging activity (60.30 %) and reducing power (absorbance 1.138) among all the samples. Fresh rose sample GC–MS analysis revealed that the there are two major compounds heptacosane 64.56 % and citronellyl propionate 28.35 %. Pentadecyl 2-phenylethyl ester oxalic acid was the second dominant compound in sun and oven-dried rose samples, 18.5 % and 14.79 % respectively

The absence of <i>Cd24</i> has no effect on dopaminergic neuronal survival or fibre innervations in the striatal 6-OHDA lesioned mouse model of Parkinson's disease.

<p>(A) Tyrosine hydroxylase (TH) immunohistochemistry on coronal sections of brain from a 6-OHDA lesioned <i>Cd24-/-</i> mouse (left column) and a <i>Cd24+/+</i> littermate (right column) at 12 days post-surgery, at the level of the striatum. (B) Representation sections of midbrain (from the respective brains provided in panel A) illustrating the loss of TH+ cells on the 6-OHDA lesioned side of the brain. (C) Optical density analysis of the TH+ fibres in the striatum indicated no difference between the genotypes. (D) Optical density analysis of the TH+ fibres in the SNpr also showed no significant difference between the genotypes. (E) Stereological estimations of the number of SNpc TH+ cells found no difference between the <i>Cd24-/-</i> and <i>Cd24+/+</i> mice. Co-ordinates in the right-hand corners of panels A indicate location of the coronal plane relative to bregma. The scale bar in panel A represents 1cm and 500μm in panels B.</p