Evaluation of the role of an antioxidant gene in NSC-34 motor neuron-like cells as a model of a motor neuron disease

Background: Spinal muscular atrophy is a rare genetic disease, which primarily affects motor neurons and predominantly occurs in children. To date, alternatives for the treatment of the disease have been controversial. Spinal muscular atrophy has a multi-factorial aetiology, with mitochondrial oxidative stress considered as the crucial pathogenic mechanism. To determine the mechanisms underlying the loss of motor neurons, NSC-34 motor neuron-like cells are often used as an in vitro model of spinal muscular atrophy. As plastin 3 (PLS3) has been demonstrated as a modifier of spinal muscular atrophy, the aim of the current study was to evaluate the neuroprotective effect of PLS3 in NSC-34 cells.  Materials and methods: Plastin 3 was overexpressed in human embryonic kidney 293T cells and NSC-34 cells via lentiviral transduction. NSC-34 cells transduced with a lentiviral vector carrying the gene for LacZ β-galactosidase served as a control. Oxidative stress was then induced by depriving cells of serum, and the protective effect of PLS3 was assessed using a cellular reactive oxygen species detection assay.  Results: While PLS3 was successfully overexpressed in human embryonic kidney 293T cells and NSC-34 cells, upregulation of this protein did not significantly decrease oxidative stress in serum-deprived NSC-34 cells relative to controls.  Conclusions: Plastin 3 overexpression in NSC-34 cells did not elicit an antioxidative effect following serum deprivation.

The effect of prolonged formalin fixation on the staining characteristics of archival human brain tissue

Background: Neurodegenerative disorders include wide range of conditions, which affect millions of people worldwide. Unfortunately, they are incurable and irreversibly progressive. Immunohistochemical staining of paraffin-fixed tissues for both diagnostic and research purposes are widely used. However, large amount of brain tissues are fixed but little is known about whether they are suitable for retrospective studies. The study aimed at investigating the effects of prolonged formalin fixation time on immunohistochemical expression of some common neurodegenerative markers in archival brain specimens.  Materials and methods: Twenty brain specimens were obtained from hu- man cadavers in the Anatomy Department of King Abdulaziz University that were prefixed in 10% formalin. They were divided into two equal groups according to time of fixation: group 1 — less than 1 year, group 2 — up to 20 years. Histological examination of white and grey matter was done using haematoxylin and eosin, luxol fast blue (LFB) for myelin staining, Con- go red for amyloid plaques, CD 68 for microglial cells, tenascin-C (large ex- tracellular matrix glycoprotein) and caspase 3 antibody for apoptotic cells.  Results: For both groups, corpus callosum sections displayed myelination with LFB staining. The distribution of CD 68 positive microglial cells was evi- dent in frontal and temporal grey matter, but not in corpus callosum sections. Strongly positive masses were seen in Congo red-stained frontal and temporal sections. Anti-caspase 3 immunostaining revealed positively stained neurons. Conclusions: Histological and immunohistochemical techniques yielded repro- ducible staining results when applied to human brain tissue stored in formalin for long periods; so they can be used in well preserved biobank material which are the most targeting research areas in neuropathology.

Oligodendrocyte pathology exceeds axonal pathology in white matter in human amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease. The majority of cases are sporadic (sALS) while the most common inherited form is due to C9orf72 mutation (C9ALS). A high burden of inclusion pathology is seen in glia (including oligodendrocytes) in ALS, especially in C9ALS. Myelin basic protein (MBP) messenger RNA (mRNA) must be transported to oligodendrocyte processes for myelination, a possible vulnerability for normal function. TDP43 is found in pathological inclusions in ALS and is a component of mRNA transport granules. Thus, TDP43 aggregation could lead to MBP loss. Additionally, the hexanucleotide expansion of mutant C9ALS, binds hnRNPA2/B1, a protein essential for mRNA transport causing potential further impairment of hnRNPA2/B1 function, and thus myelination. Using immunohistochemistry p62 and TDP43 in human post mortem tissue, we found a high burden of glial inclusions in the prefrontal cortex, precentral gyrus and spinal cord in ALS, which was greater in C9ALS than sALS cases. Double staining demonstrated the majority of these inclusions were in oligodendrocytes. Using immunoblotting, we demonstrated reduced MBP protein levels relative to PLP (a myelin component that relies on protein not mRNA transport) and neurofilament protein (an axonal marker) in the spinal cord. This MBP loss was disproportionate to the level of PLP and axonal loss, suggesting that impaired mRNA transport may be partly responsible. Finally, we show that in C9ALS cases, the level of oligodendroglial inclusions correlates inversely with levels of hnRNPA2/B1 and the number of oligodendrocyte precursor cells. We conclude that there is considerable oligodendrocyte pathology in ALS, which at least partially reflects impairment of mRNA transport

Genetic modifiers ameliorate endocytic and neuromuscular defects in a model of spinal muscular atrophy

© 2020 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/), which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made.Background: Understanding the genetic modifiers of neurodegenerative diseases can provide insight into the mechanisms underlying these disorders. Here, we examine the relationship between the motor neuron disease spinal muscular atrophy (SMA), which is caused by reduced levels of the survival of motor neuron (SMN) protein, and the actin-bundling protein Plastin 3 (PLS3). Increased PLS3 levels suppress symptoms in a subset of SMA patients and ameliorate defects in SMA disease models, but the functional connection between PLS3 and SMN is poorly understood.Results: We provide immunohistochemical and biochemical evidence for large protein complexes localized in vertebrate motor neuron processes that contain PLS3, SMN and members of the hnRNP F/H family of proteins. Using a Caenorhabditis elegans (C. elegans) SMA model, we determine that overexpression of PLS3 or loss of the C. elegans hnRNP F/H ortholog SYM-2 enhances endocytic function and ameliorates neuromuscular defects caused by decreased SMN-1 levels. Furthermore, either increasing PLS3 or decreasing SYM-2 levels suppresses defects in a C. elegans ALS model.Conclusions: We propose that hnRNP F/H act in the same protein complex as PLS3 and SMN and that the function of this complex is critical for endocytic pathways, suggesting that hnRNP F/H proteins could be potential targets for therapy development.Peer reviewe

Efficacy and safety of single pill combination of amlodipine and valsartan in hypertensive Saudi patients

OBJECTIVE: A significant global health issue that affects 25.5% of Saudi people is hypertension (HTN). According to international recommendations, most HTN patients require more than one therapy to reach their blood pressure targets (BP). Therefore, it would be preferable to utilize two medications from distinct classes separately or in a predetermined combination. According to recent studies, a single-pill combination (SPC) may be more efficient. This study evaluated the safety and tolerability of Amlodipine/Valsartan (Aml/Val) SPC in Saudi hypertensive patients, as well as the effectiveness of the medication. PATIENTS AND METHODS: Observational research was done prospectively at the King Fahad Armed Forces Hospital in Jeddah, Saudi Arabia. The effectiveness of the treatment and the percentage of 159 hypertensive patients who achieved the target blood pressure values (140/90; 130/80 mmHg) among those with diabetes mellitus (DM), chronic kidney disease (CKD), other cardiovascular disorders, and responders were assessed from the beginning to the endpoint (week 23). RESULTS: According to the results, taking Aml/Val SPC significantly lowered all patients’ baseline systolic and diastolic blood pressure readings by -17.97 and 8.58 mmHg, respectively. 43.4% of patients successfully met their BP therapeutic objectives by bringing their blood pressure levels back to normal, including 51.4% of patients under 65, 39.3% of patients with chronic kidney disease, and 26.2% of diabetic patients. Aml/Val 10/160 mg significantly lowers SBP, more than Aml/Val 5/160 mg (-13.32% vs. -9.00%, p<0.050). Vertigo (6.30%), respiratory tract infections (4.0%), and ankle edema (2.50%) were the most frequent adverse events. CONCLUSIONS: Aml/Val SPC therapy effectively lowered BP and had few side effects while being well-tolerated in people with hypertension

Neuroprotective Effect of Red Sea Marine Sponge <i>Xestospongia testudinaria</i> Extract Using In Vitro and In Vivo Diabetic Peripheral Neuropathy Models

Diabetic peripheral neuropathy (DPN) is a common complication of diabetes. Oxidative stress plays an important role in the pathophysiology of DPN. Red Sea marine sponge Xestospongia testudinaria extract has a promising neuroprotective effect, presumably owing to its antioxidant and anti-inflammatory properties. Thus, this study aimed to investigate the neuroprotective effect of the sponge X. testudinaria extract on in vitro and in vivo models of DPN. Mice dorsal root ganglia (DRG) were cultured with high glucose (HG) media and used as an in vitro model of DPN. Some of the DRGs were pre-treated with 2 mg/mL of X. testudinaria. The X. testudinaria extract significantly improved the HG-induced decreased neuronal viability and the neurite length. It improved the oxidative stress biomarkers in DRG cultures. The DPN model was induced in vivo by an injection of streptozotocin at a dose of 150 mg/kg in mice. After 35 days, 0.75 mg/kg of the X. testudinaria extract improved the hot hyperalgesia and the DRG histology. Although the sponge extract did not reduce hyperglycemia, it ameliorated the oxidative stress markers and pro-inflammatory markers in the DRG. In conclusion, the current study demonstrates the neuroprotective effect of Red Sea sponge X. testudinaria extract against experimentally induced DPN through its antioxidant and anti-inflammatory mechanisms

The combined effect of honey and olive oil against methotrexate mediated hepatotoxicity in rats: A biochemical, histological and immunohistological study

Background. Honey and olive oil are natural products that have high nutritional values, and therapeutic properties. Cytotoxic drugs, like methotrexate (MTX) are used to treat malignancies in tumour cells; however, these drugs also have serious side effects that could threaten the patient's life. Aim. To evaluate the potential protective effects of honey and olive oil, administered alone or together, against MTX-induced hepatotoxicity in rats. Methods. Adult male albino rats were divided: Group I: negative control (n=8); II: honey (daily by oral 1.2 g/kg bwt (n=8), III: olive oil (1 ml/day) (n=8), IV: single intraperitoneal injection of MTX (20 mg/kg bwt) (n=8), V: diluted honey for 3 days before injection of MTX (n=8), Group VI: olive oil for 3 days before injection of MTX (n=8), Group VII: both honey and olive oil for 3 days before injection of MTX (n=8). After treatment, rats were sacrified and blood samples were collected to determine liver function parameters, liver tissue used to measure the oxidative (malondialdehyde), antioxidative parameters (superoxide dismutase, catalase and glutathione peroxidase), histological and immunohistochemical techniques. Results. The administration of honey and olive oil exerted a protective effect against MTX-induced hepatotoxicity, as demonstrated by the normalization of the liver enzymes, proteins and total bilirubin and by the histopathological and immunohistological changes observed in the livers. Both agents also reversed the oxidative damage in the liver by decreasing level of MDA levels and increasing the antioxidant related by enzymes in the liver homogenates compared to the control rats. These effects were more evident when the two agents were administered together. Conclusion. The combined intake of honey and olive oil could be hepatoprotective. Co-administration of these agents might form an effective adjuvant therapy and minimize side effects of chemoherapy in cancerous patients