Native α-Synuclein, 3-Nitrotyrosine Proteins, and Patterns of Nitro-α-Synuclein-Immunoreactive Inclusions in Saliva and Submandibulary Gland in Parkinson's Disease

Native α-Synuclein, 3-Nitrotyrosine Proteins, and Patterns of Nitro-α-Synuclein-Immunoreactive Inclusions in Saliva and Submandibulary Gland in Parkinson’s Disease

Background. Salivary α-synuclein (aSyn) and its nitrated form, or 3-nitrotyrosine-α-synuclein (3-NT-αSyn), hold promise as biomarkers for idiopathic Parkinson’s disease (IPD). Nitrative stress that is characterized by an excess of 3-nitrotyrosine proteins (3-NT-proteins) has been proposed as a pathogenic mechanism in IPD.
The objective is to study the pathological role of native αSyn, 3-NT-αSyn, and 3-NT-proteins in the saliva and submandibulary glands of patients with IPD. Methods. The salivary and serum αSyn and 3-NT-proteins concentration is evaluated with ELISA in patients and controls. Correlations of αSyn and 3-NT-proteins content with clinical features of the disease are examined.
Immunohistochemical 3-NT-αSyn expression in submandibulary gland sections is analyzed. Results. (a) Salivary concentration and saliva/serum ratios of native αSyn and 3-NT-proteins are similar in patients and controls; (b) salivary αSyn and 3-NT-proteins do not correlate with any clinical feature; and (c) three patterns of 3-NT-αSyn-positive inclusions are observed on histological sections: rounded “Lewy-type” aggregates of 10-25 µm in diameter, coarse deposits with varied morphology, and spheroid inclusions or bodies of 3-5 µm in diameter.
“Lewy-type” and coarse inclusions are observed in the interlobular connective tissue of the gland, and small-sized bodies are located within the cytoplasm of duct cells. “Lewy-type” inclusions are only observed in patients, and the remaining patterns of inclusions are observed in both the patients and controls. Conclusions.
The patients’ saliva presents a similar concentration of native αSyn and 3-nitrotyrosine-proteins than that of the controls, and no correlations with clinical features are found. These findings preclude the utility of native αSyn in the saliva as a biomarker, and they indicate the absence of nitrative stress in the saliva and serum of patients.
As regards nitrated αSyn, “Lewy-type” inclusions expressing 3-NT-αSyn are observed in the patients, not the controls-a novel finding that suggests that a biopsy of the submandibulary gland, if proven safe, could be a useful technique for diagnosing IPD.
Finally, to our knowledge, this is also the first description of 3-NT-αSyn-immunoreactive intracytoplasmic bodies in cells that are located outside the nervous system. These intracytoplasmic bodies are present in duct cells of submandibulary gland sections from all subjects regardless of their pathology, and they can represent an aging or involutional change. Further immunostaining studies with different antibodies and larger samples are needed to validate the data.

A Method for Analysis of Nitrotyrosine-Containing Proteins by Immunoblotting Coupled with Mass Spectrometry

Nitrotyrosine formation is caused by presence of reactive oxygen and nitrogen species. Nitration is a very selective process leading to specific modification of only a few tyrosines in protein molecule. 2D electrophoresis and western blotting techniques coupled with mass spectrometry are common methods used in analysis of proteome.
Here we describe protocol for analysis of peroxynitrite-induced protein nitration in isolated mitochondria. Mitochondrial proteins are separated by 2D electrophoresis and transferred to nitrocellulose membrane. Membranes are then incubated with antibodies against nitrotyrosine. Positive spots are compared with corresponding Coomassie-stained gels, and protein nitration is confirmed with mass spectrometry techniques.

Muscle Cortisol Levels, Expression of Glucocorticoid Receptor and Oxidative Stress Markers in the Teleost Fish Argyrosomus regius Exposed to Transport Stress

Fish commercial transport is an ordinary practice in the aquaculture industry. This study aimed to investigate the effect of a 48 h transport stress on stress response of meagre (Argyrosomus regius) juveniles. Radioimmunoassay (RIA) and Real-Time PCR were used to evaluate muscle cortisol levels and to assess glucocorticoid receptor (gr) gene expression in fish muscle and liver, respectively.
Presence and localization of various oxidative stress markers were investigated in different tissues by immunohistochemistry. A significant increase in muscle cortisol levels was observed after loading but a significant decrease occurred after 16 h from departure even without returning to control levels.
Native α-Synuclein, 3-Nitrotyrosine Proteins, and Patterns of Nitro-α-Synuclein-Immunoreactive Inclusions in Saliva and Submandibulary Gland in Parkinson's Disease
Molecular analysis on stress response revealed an increase in muscle gr expression after fish loading that started decreasing during the travel returning to the control level at the end of the transport. Instead, no differences in liver gr expression were observed along the different sampling points. Immunostaining for heat shock protein 70 (HSP70), 4-hydroxy-2-nonenal (HNE), nitrotyrosine (NT) and 8-hydroxy-2′-deoxyguanosine (8-OHdG) antibodies was detected in several organs.
Notably, a higher NT immunostaining intensity was evident in skin and gills of the transported animals with respect to controls. Results demonstrated that cortisol and gr are useful indicators of stressful conditions in transported fish.

Inhibition of Cochlear HMGB1 Expression Attenuates Oxidative Stress and Inflammation in an Experimental Murine Model of Noise-Induced Hearing Loss

Noise-induced hearing loss (NIHL) is a common inner ear disease but has complex pathological mechanisms, one of which is increased oxidative stress in the cochlea. The high-mobility group box 1 (HMGB1) protein acts as an inflammatory mediator and shows different activities with redox modifications linked to the generation of reactive oxygen species (ROS).
We aimed to investigate whether manipulation of cochlear HMGB1 during noise exposure could prevent noise-induced oxidative stress and hearing loss. Sixty CBA/CaJ mice were divided into two groups. An intraperitoneal injection of anti-HMGB1 antibodies was administered to the experimental group; the control group was injected with saline.
Thirty minutes later, all mice were subjected to white noise exposure. Subsequent cochlear damage, including auditory threshold shifts, hair cell loss, expression of cochlear HMGB1, and free radical activity, was then evaluated. The levels of HMGB1 and 4-hydroxynonenal (4-HNE), as respective markers of reactive nitrogen species (RNS) and ROS formation, showed slight increases on post-exposure day 1 and achieved their highest levels on post-exposure day 4.
After noise exposure, the antibody-treated mice showed markedly less ROS formation and lower expression of NADPH oxidase 4 (NOX4), nitrotyrosine, inducible nitric oxide synthase (iNOS), and intercellular adhesion molecule-1 (ICAM-1) than the saline-treated control mice.

Nitrotyrosine Antibody

abx441444-100ug 100 ug
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abx441725-100ug 100 ug
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Nitrotyrosine Antibody

abx442006-100ug 100 ug
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abx442287-100ug 100 ug
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Nitrotyrosine Antibody

abx442568-100ug 100 ug
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Nitrotyrosine Antibody

abx442849-100ug 100 ug
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Nitrotyrosine Antibody

abx443129-100ug 100 ug
EUR 693.6

Nitrotyrosine Antibody

abx443409-100ug 100 ug
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Nitrotyrosine Antibody

abx443690-100ug 100 ug
EUR 727.2

Nitrotyrosine Antibody

abx443971-100ug 100 ug
EUR 710.4

Nitrotyrosine Antibody

abx444252-100ug 100 ug
EUR 710.4

Nitrotyrosine Antibody

abx444533-100ug 100 ug
EUR 710.4

Nitrotyrosine Antibody

20-abx444883
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Nitrotyrosine Antibody

5416-50 each
EUR 405.6

Nitrotyrosine Antibody

abx140246-01mg 0.1 mg
EUR 644.4

Nitrotyrosine Antibody

abx023270-1ml 1 ml
EUR 811.2

Nitrotyrosine Antibody

GWB-A3CF01 0.1 mg Ask for price

Nitrotyrosine Antibody

GWB-63CB0B 1 ml Ask for price

NITROTYROSINE, Antibody

GWB-E886B9 0.1 mg Ask for price

Nitrotyrosine, Antibody

GWB-931C05 0.1 mg Ask for price

Nitrotyrosine Antibody

abx140246-100g 100 µg
EUR 450

Nitrotyrosine Antibody

abx444883-100l 100 µl
EUR 362.5

Nitrotyrosine (NT) Antibody

20-abx100841
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Nitrotyrosine (NT) Antibody

abx100841-100l 100 µl
EUR 287.5

Nitrotyrosine (NT) Antibody

abx100841-1ml 1 ml
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Nitrotyrosine (NT) Antibody

abx100841-200l 200 µl
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Nitrotyrosine Antibody (PE)

abx444252-100g 100 µg
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Nitrotyrosine Antibody (7A5)

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EUR 392.4

Nitrotyrosine Antibody (APC)

abx442568-100l 100 µl
EUR 625

Nitrotyrosine Antibody (HRP)

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EUR 612.5

Nitrotyrosine Antibody (HM11)

5414-50 each
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Nitrotyrosine Antibody (FITC)

abx443129-100g 100 µg
EUR 612.5

Nitrotyrosine Antibody (PerCP)

abx443971-100g 100 µg
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Nitrotyrosine Antibody (Biotin)

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Nitrotyrosine Antibody (ATTO488)

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Nitrotyrosine Antibody, Clone 39B6

SMC-154C 0.025mg
EUR 167
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is not conjugated.

Nitrotyrosine Antibody, Clone 39B6

SMC-154D 0.1mg
EUR 343
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is not conjugated.

Nitrotyrosine Antibody, Clone 39B6

SMC-154S 0.012mg
EUR 44
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine | Monkey | Dog | Rabbit Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is not conjugated.

Nitrotyrosine Antibody, Clone 39B6: APC

SMC-154D-APC 0.1mg
EUR 395
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine | Monkey | Dog | Rabbit Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with APC.

Nitrotyrosine Antibody, Clone 39B6: HRP

SMC-154D-HRP 0.1mg
EUR 383
Description: A monoclonal antibody from clone 39B6 against Human | Mouse | Rat | Bovine | Monkey | Dog | Rabbit Nitrotyrosine. The host species for the production of this antibody is Mouse. The antigen used for immunization is 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin. The antibody is tested and validated for WB, IHC, ICC/IF, IP, ELISA, FCM, AM assays with the following recommended dilutions: WB (1:1400), IHC (1:100). This MAb for Nitrotyrosine is conjugated with HRP.
A significant amelioration was also observed in the threshold shifts of the auditory brainstem response and the loss of outer hair cells in the antibody-treated versus the saline-treated mice. Our results suggest that inhibition of HMGB1 by neutralization with anti-HMGB1 antibodies prior to noise exposure effectively attenuated oxidative stress and subsequent inflammation. This procedure could therefore have potential as a therapy for NIHL.

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