Publications by authors named "Sikha Saha"

25 Publications

  • Page 1 of 1

Barium yttrium fluoride based upconversion nanoparticles as dual mode image contrast agents.

Mater Sci Eng C Mater Biol Appl 2021 May 6;124:111937. Epub 2021 Feb 6.

School of Chemical and Process Engineering, Faculty of Engineering and Physical Sciences, University of Leeds, Leeds LS2 9JT, United Kingdom.

Dual labeled contrast agents could provide better complementary information for bioimaging than available solely from a single modality. In this paper we investigate the suitability of Yb and Er-doped BaYF upconversion nanoparticles (UCNPs) as both optical and X-ray micro computed tomography (μCT) contrast agents. Stable, aqueous UCNP dispersions were synthesised using a hydrothermal method with the addition of polyethyleneimine (PEI). UCNPs were single crystal and had a truncated cuboidal and/or truncated octahedral morphology, with average particle size of 47 ±9 nm from transmission electron microscopy which was further used to characterize the structure and composition in detail. A zeta potential value of +51 mV was measured for the aqueous nanoparticle dispersions which is beneficial for cell permeability. The outer hydrated PEI layer is also advantageous for the attachment of proteins for targeted delivery in biological systems. The prepared UCNPs were proven to be non-toxic to endothelial cells up to a concentration of 3.5 mg/mL, when assessed using an MTT assay. The particles showed intense green upconversion photoluminescence when excited at a wavelength of 976 nm using a diode laser. Quantitative X-ray μCT contrast imaging confirmed the potential of these UCNPs as X-ray contrast agents and confirming their dual modality for bioimaging.
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http://dx.doi.org/10.1016/j.msec.2021.111937DOI Listing
May 2021

Affimer-Based Europium Chelates Allow Sensitive Optical Biosensing in a Range of Human Disease Biomarkers.

Sensors (Basel) 2021 Jan 27;21(3). Epub 2021 Jan 27.

Bionanotechnology Group, School of Biomedical Science, University of Leeds, Leeds LS2 9JT, UK.

The protein biomarker measurement has been well-established using ELISA (enzyme-linked immunosorbent assay), which offers good sensitivity and specificity, but remains slow and expensive. Certain clinical conditions, where rapid measurement or immediate confirmation of a biomarker is paramount for treatment, necessitate more rapid analysis. Biosensors offer the prospect of reagent-less, processing-free measurements at the patient's bedside. Here, we report a platform for biosensing based on chelated Eu against a range of proteins including biomarkers of cardiac injury (human myoglobin), stroke (glial fibrillary acidic protein (GFAP)), inflammation (C-reactive protein (CRP)) and colorectal cancer (carcinoembryonic antigen (CEA)). The Eu ions are chelated by modified synthetic binding proteins (Affimers), which offer an alternative targeting strategy to existing antibodies. The fluorescence characteristics of the Eu complex with modified Affimers against human myoglobin, GFAP, CRP and CEA were measured in human serum using λ = 395 nm, λ = 590 and 615 nm. The Eu-Affimer based complex allowed sensitive detection of human myoglobin, GFAP, CRP and CEA proteins as low as 100 fM in (100-fold) diluted human serum samples. The unique dependence on Eu fluorescence in the visible region (590 and 615 nm) was exploited in this study to allow rapid measurement of the analyte concentration, with measurements in 2 to 3 min. These data demonstrate that the Affimer based Eu complexes can function as nanobiosensors with potential analytical and diagnostic applications.
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http://dx.doi.org/10.3390/s21030831DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7865513PMC
January 2021

Editorial: Cerebrovascular and Neurodegenerative Diseases - New Insights Into Molecular Cell Biology and Therapeutic Targets.

Front Neurol 2019 13;10:1322. Epub 2019 Dec 13.

School of Pharmacy and Medical Sciences, University of Bradford, Bradford, United Kingdom.

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http://dx.doi.org/10.3389/fneur.2019.01322DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6923269PMC
December 2019

Subchronic hypoxia pretreatment on brain pathophysiology in unilateral common carotid artery occluded albino rats.

Indian J Pharmacol 2018 Jul-Aug;50(4):185-191

Division of Cardiovascular and Diabetes Research, Leeds Institute of Cardiovascular and Metabolic Medicine, University of Leeds, Leeds, UK.

Objective: This study was aimed to assess the effect of unilateral common carotid artery occlusion on brain pathophysiology in rats pretreated with subchronic hypoxia.

Materials And Methods: Rats (200 ± 20 g) were randomized into three groups: Group 1 served as sham, Group 2 were normoxic (21% O and 79% N), and Group 3 were hypoxia preconditioned (10% O and 90% N) for 21 days before left common carotid artery occlusion (LCCAO). The LCCAO was done for 75 min followed by reperfusion for 12 h. Neurological scores were recorded. Serum malondialdehyde (MDA) and nitric oxide (NO) levels at pre- and 12 h post-LCCAO were measured. Brain histopathological assessments were also done.

Results: Higher neurological deficits scores in Group 2 as compared to Group 3 rats were noticed. Serum MDA and NO levels at 12 h post-LCCAO in Group 2 rats showed significant elevation as compared to preocclusion levels. Group 3 rats did not show such elevations. On histopathology of left and right cerebral hemispheres of Group 1 (sham) did not show any specific changes. In Group 2 rats, the right cerebral hemisphere (nonoccluded) showed no areas of ischemia-induced brain changes, but in the left side (occlusive), there were features of ischemic brain damage including cerebral edema. In the case of Group 3 rats, there were less ischemic damages in the left occluded side as compared to the left side of the Group 2 rats.

Conclusion: This study clearly demonstrates that subchronic hypoxia pretreatment can reduce ischemic brain injury by unilateral common carotid artery occlusion in rats.
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http://dx.doi.org/10.4103/ijp.IJP_312_17DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6234715PMC
March 2019

Selective cellular imaging with lanthanide-based upconversion nanoparticles.

J Biophotonics 2019 04 2;12(4):e201800256. Epub 2019 Jan 2.

Leeds Institute for Cardiovascular and Metabolic Medicine (LICAMM), Faculty of Medicine and Health, University of Leeds, Leeds LS2 9JT, UK.

Upconversion nanoparticles (UCNPs) with sodium yttrium fluoride, NaYF (host lattice) doped with Yb (sensitizer) and Er (activator) were synthesized via hydrothermal route incorporating polyethyleneimine (PEI) for their long-term stability in water. The cationic PEI-modified UCNPs with diameter 20 ± 4 nm showed a zeta potential value of +36.5 mV and showed an intense, visible red luminescence and low-intensity green emission with 976 nm laser excitation. The particles proven to be nontoxic to endothelial cells, with a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay, showing 90% to 100% cell viability, across a wide range of UCNP concentrations (0.3 ng/mL-0.3 mg/mL) were used in multiphoton imaging. Multiphoton cellular imaging and emission spectroscopy data reported here prove that the UCNPs dispersed in cell culture media are predominantly concentrated in the cytoplasm than the cell nucleus. The energy transfer from PEI-coated UCNPs to surrounding media for red luminescence in the biological system is also highlighted with spectroscopic measurements. Results of this study propose that UCNPs can, therefore, be used for cytoplasm selective imaging together with multiphoton dyes (eg, 4',6-diamidino-2-phenylindole (DAPI)) that are selective to cell nucleus.
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http://dx.doi.org/10.1002/jbio.201800256DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7065621PMC
April 2019

A novel dynamic multicellular co-culture system for studying individual blood-brain barrier cell types in brain diseases and cytotoxicity testing.

Sci Rep 2018 06 8;8(1):8784. Epub 2018 Jun 8.

Leeds Institute of Cardiovascular and Metabolic Medicine, Faculty of Medicine and Health, University of Leeds, Leeds, UK.

Blood brain barrier (BBB) cells play key roles in the physiology and pathology of the central nervous system (CNS). BBB dysfunction is implicated in many neurodegenerative diseases, including Alzheimer's disease (AD). The BBB consists of capillary endothelial cells, pericytes encircling the endothelium and surrounding astrocytes extending their processes towards it. Although there have been many attempts to develop in vitro BBB models, the complex interaction between these cell types makes it extremely difficult to determine their individual contribution to neurotoxicity in vivo. Thus, we developed and optimised an in vitro multicellular co-culture model within the Kirkstall Quasi Vivo System. The main aim was to determine the optimal environment to culture human brain primary endothelial cells, pericytes and astrocytes whilst maintaining cellular communication without formation of a barrier in order to assess the contribution of each cell type to the overall response. As a proof of concept for the present system, the effects of amyloid-beta 25-35 peptide (Aβ25-35), a hallmark of AD, were explored. This multicellular system will be a valuable tool for future studies on the specific roles of individual BBB cell type (while making connection with each other through medium) in CNS disorders as well as in cytotoxicity tests.
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http://dx.doi.org/10.1038/s41598-018-26480-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5993789PMC
June 2018

Tau pathology and neurochemical changes associated with memory dysfunction in an optimised murine model of global cerebral ischaemia - A potential model for vascular dementia?

Neurochem Int 2018 09 10;118:134-144. Epub 2018 Apr 10.

Discovery and Translational Science Department, Leeds Institute of Cardiovascular and Metabolic Medicine, University of Leeds, Leeds, LS2 9JT, UK. Electronic address:

Cerebral ischemia is known to be a major cause of death and the later development of Alzheimer's disease and vascular dementia. However, ischemia induced cellular damage that initiates these diseases remain poorly understood. This is primarily due to lack of clinically relevant models that are highly reproducible. Here, we have optimised a murine model of global cerebral ischaemia with multiple markers to determine brain pathology, neurochemistry and correlated memory deficits in these animals. Cerebral ischaemia in mice was induced by bilateral common carotid artery occlusion. Following reperfusion, the mice were either fixed with 4% paraformaldehyde or decapitated under anaesthesia. Brains were processed for Western blotting or immunohistochemistry for glial (GLT1) and vesicular (VGLUT1, VGLUT2) glutamate transporters and paired helical filament (PHF1) tau. The PHF1 tau is the main component of neurofibrillary tangle, which is the pathological hallmark of Alzheimer's disease and vascular dementia. The novel object recognition behavioural assay was used to investigate the functional cognitive consequences in these mice. The results show consistent and selective neuronal and glial cell changes in the hippocampus and the cortex together with significant reductions in GLT1 (***P < 0.001), VGLUT1 (**P < 0.01) and VGLUT2 (***P < 0.001) expressions in the hippocampus in occluded mice as compared to sham-operated animals. These changes are associated with increased PHF1 (***P < 0.0001) protein and a significant impairment of performance (*p < 0.0006, N = 6/group) in the novel object recognition test. This model represents a useful tool for investigating cellular, biochemical and molecular mechanisms of global cerebral ischaemia and may be an ideal preclinical model for vascular dementia.
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http://dx.doi.org/10.1016/j.neuint.2018.04.004DOI Listing
September 2018

The theoretical molecular weight of NaYF :RE upconversion nanoparticles.

Sci Rep 2018 01 18;8(1):1106. Epub 2018 Jan 18.

School of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds, Leeds, LS2 9JT, United Kingdom.

Upconversion nanoparticles (UCNPs) are utilized extensively for biomedical imaging, sensing, and therapeutic applications, yet the molecular weight of UCNPs has not previously been reported. Herein, we present a theory based upon the crystal structure of UCNPs to estimate the molecular weight of UCNPs: enabling insight into UCNP molecular weight for the first time. We estimate the theoretical molecular weight of various UCNPs reported in the literature, predicting that spherical NaYF4 UCNPs ~ 10 nm in diameter will be ~1 MDa (i.e. 10 g/mol), whereas UCNPs ~ 45 nm in diameter will be ~100 MDa (i.e. 10 g/mol). We also predict that hexagonal crystal phase UCNPs will be of greater molecular weight than cubic crystal phase UCNPs. Additionally we find that a Gaussian UCNP diameter distribution will correspond to a lognormal UCNP molecular weight distribution. Our approach could potentially be generalised to predict the molecular weight of other arbitrary crystalline nanoparticles: as such, we provide stand-alone graphic user interfaces to calculate the molecular weight both UCNPs and arbitrary crystalline nanoparticles. We expect knowledge of UCNP molecular weight to be of wide utility in biomedical applications where reporting UCNP quantity in absolute numbers or molarity will be beneficial for inter-study comparison and repeatability.
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http://dx.doi.org/10.1038/s41598-018-19415-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5773537PMC
January 2018

Photoluminescence intensity ratio of Eu-conjugated lactates-A simple optical imaging technique for biomarker analysis for critical diseases.

J Biophotonics 2018 05 18;11(5):e201700199. Epub 2017 Dec 18.

Leeds Cardiovascular and Metabolic Medicine, Faculty of Medicine and Health, University of Leeds, Leeds, UK.

Instant measurement of elevated biomarkers such as lactic acid offers the most promising approaches for early treatment and prevention of many critical diseases including cardiac arrest, stroke, septic shock, trauma, liver dysfunction, as well as for monitoring lactic acid level during intense exercise. In the present study, a unique dependence of visible photoluminescence of Eu ions resulting from D to F transitions, which can be exploited for rapid detection of biomarkers, both in vitro and ex vivo, has been reported. It is observed that the integrated intensity ratio of photoluminescence signals dominating at 591 and 616 nm originating from D to F and D to F transitions in Eu ions can be used as a biosensing and bioimaging tool for detection of biomarkers released at disease states. The Eu integrated photoluminescence intensity ratio approach reported herein for optical detection of lactates in blood serum, plasma and confocal imaging of brain tissues has very high potential for exploitation of this technique in both in vitro monitoring and in vivo bioimaging applications for the detection of biomarkers in various diseases states.
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http://dx.doi.org/10.1002/jbio.201700199DOI Listing
May 2018

α-tocopherol supplementation prevents lead acetate and hypoxia-induced hepatic dysfunction.

Indian J Pharmacol 2015 May-Jun;47(3):285-91

Defence Institute of Physiology and Allied Sciences, DRDO, Timarpur, New Delhi, India.

Objective: Lead (Pb) is a long-known poison of environment and industrial origin. Its prolonged exposure affects cellular material and alters cellular genetics and produces oxidative damages. In this study, we investigated the exposure of chronic sustained hypoxia or lead acetate alone or in combination with or without supplementation of α-tocopherol on hepatic oxidative and nitrosative stress in rats.

Materials And Methods: The rats weighing 165 ± 5 g were exposed to chronic sustained hypoxia (10% oxygen) or lead acetate (25 mg/kg of body weight, intraperitoneally) alone or in combination with or without supplementation of α-tocopherol (10 mg/100 g b.wt, intramuscularly). The body weight of all the rats was recorded on the day 1 of the treatment and the day of sacrifice. Serum lipid profile was estimated by using a biochemical analyzer. Oxidant and enzymatic antioxidants status was evaluated by using spectrophotometer. Serum levels of hypoxia inducible factor-1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF) were measured by using ELISA technique. Histopathological assessments of hepatic tissue were also done.

Results: Exposure of both lead and hypoxia showed decreased body weight, altered serum lipid profile, oxidant and enzymatic antioxidants status, serum HIF-1α and VEGF concentrations. Simultaneous α-tocopherol supplementation showed beneficial effects to all these alterations. Histopathological observations also showed hepatic degenerative changes after lead or hypoxia exposure either alone or in combination, but remarkable improvement has been noticed after α-tocopherol supplementation.

Conclusion: Supplementation of α-tocopherol is beneficial to counter both lead acetate and hypoxia induced hepatic cytotoxicities possibly by reducing oxidative and nitrosative stress.
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http://dx.doi.org/10.4103/0253-7613.157126DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4450554PMC
May 2016

Therapeutic benefits of nanoparticles in stroke.

Front Neurosci 2015 19;9:182. Epub 2015 May 19.

Division of Cardiovascular and Diabetes Research, Leeds Institute of Cardiovascular and Metabolic Medicine, University of Leeds Leeds, UK.

Stroke represents one of the major causes of death and disability worldwide, for which no effective treatments are available. The thrombolytic drug alteplase (tissue plasminogen activator or tPA) is the only treatment for acute ischemic stroke but its use is limited by several factors including short therapeutic window, selective efficacy, and subsequent haemorrhagic complications. Numerous preclinical studies have reported very promising results using neuroprotective agents but they have failed at clinical trials because of either safety issues or lack of efficacy. The delivery of many potentially therapeutic neuroprotectants and diagnostic compounds to the brain is restricted by the blood-brain barrier (BBB). Nanoparticles (NPs), which can readily cross the BBB without compromising its integrity, have immense applications in the treatment of ischemic stroke. In this review, potential uses of NPs will be summarized for the treatment of ischemic stroke. Additionally, an overview of targeted NPs will be provided, which could be used in the diagnosis of stroke. Finally, the potential limitations of using NPs in medical applications will be mentioned. Since the use of NPs in stroke therapy is now emerging and is still in development, this review is far from comprehensive or conclusive. Instead, examples of NPs and their current use will be provided, as well as the potentials of NPs in an effort to meet the high demand of new therapies in stroke.
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http://dx.doi.org/10.3389/fnins.2015.00182DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4436818PMC
June 2015

Centrally administered CDP-choline induced cardiovascular responses are mediated by activation of the central phospholipase-prostaglandin signaling cascade.

Brain Res 2014 May 2;1563:61-71. Epub 2014 Apr 2.

Department of Physiology, Faculty of Veterinary Medicine, Uludag University, Bursa 16059, Turkey. Electronic address:

The present study was designed to determine the involvement of central prostaglandin synthesis on the pressor and bradycardic effect of cytidine 5'-diphosphocholine (CDP-choline). Intracerebroventricular (i.c.v.) administration of CDP-choline was made and blood pressure and heart rate were recorded in male Sprague Dawley rats throughout this study. Microdialysis and immunohistochemical studies were performed to measure extracellular total prostaglandin concentration and to show cyclooxygenase-1 and -2 (COX-1 and -2) immunoreactivities, respectively, in the posterior hypothalamic area. Moreover, rats were pretreated (i.c.v) with mepacrine [a phospholipase A2 (PLA2) inhibitor], ibuprofen [a nonselective COX inhibitor], neomycine [a phospholipase C (PLC) inhibitor] or furegrelate [a thromboxane A2 (TXA2) synthesis inhibitor] 5 min prior to the injection of CDP-choline to determine the effects of these inhibitors on cardiovascular responses to CDP-choline. Control rats were pretreated (i.c.v) with saline. CDP-choline caused a dose- and time-dependent increase in blood pressure and decrease in heart rate. Immunohistochemical studies showed that CDP-choline increased COX-1 and -2 immunoreactivities in the posterior hypothalamic area. CDP-choline also elevated hypothalamic extracellular total prostaglandin concentration by 62%, as shown in microdialysis studies. Mepacrine or ibuprofen pretreatments almost completely blocked the pressor and bradycardic responses to CDP-choline while neomycine or furegrelate partially attenuated the drug-induced cardiovascular effects. The results suggest that CDP-choline may stimulate prostaglandin synthesis through the activation of PLA2, cyclooxygenases (COX-1 and -2) and prostaglandins and at least TXA2, may mediate the drug׳s cardiovascular effects.
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http://dx.doi.org/10.1016/j.brainres.2014.03.040DOI Listing
May 2014

Nanotechnology for the detection and therapy of stroke.

Adv Healthc Mater 2014 Nov 1;3(11):1703-20. Epub 2014 Apr 1.

School of Medicine, University of Leeds, Leeds, LS2 9JT, UK.

Over the years, nanotechnology has greatly developed, moving from careful design strategies and synthesis of novel nanostructures to producing them for specific medical and biological applications. The use of nanotechnology in diagnostics, drug delivery, and tissue engineering holds great promise for the treatment of stroke in the future. Nanoparticles are employed to monitor grafted cells upon implantation, or to enhance the imagery of the tissue, which is coupled with a noninvasive imaging modality such as magnetic resonance imaging, computed axial tomography or positron emission tomography scan. Contrast imaging agents used can range from iron oxide, perfluorocarbon, cerium oxide or platinum nanoparticles to quantum dots. The use of nanomaterial scaffolds for neuroregeneration is another area of nanomedicine, which involves the creation of an extracellular matrix mimic that not only serves as a structural support but promotes neuronal growth, inhibits glial differentiation, and controls hemostasis. Promisingly, carbon nanotubes can act as scaffolds for stem cell therapy and functionalizing these scaffolds may enhance their therapeutic potential for treatment of stroke. This Progress Report highlights the recent developments in nanotechnology for the detection and therapy of stroke. Recent advances in the use of nanomaterials as tissue engineering scaffolds for neuroregeneration will also be discussed.
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http://dx.doi.org/10.1002/adhm.201400009DOI Listing
November 2014

The effect of centrally injected CDP-choline on respiratory system; involvement of phospholipase to thromboxane signaling pathway.

Respir Physiol Neurobiol 2014 May 18;195:50-8. Epub 2014 Feb 18.

Department of Physiology, Faculty of Veterinary Medicine, Uludag University, Bursa 16059, Turkey. Electronic address:

CDP-choline is an endogenous metabolite in phosphatidylcholine biosynthesis. Exogenous administration of CDP-choline has been shown to affect brain metabolism and to exhibit cardiovascular, neuroendocrine neuroprotective actions. On the other hand, little is known regarding its respiratory actions and/or central mechanism of its respiratory effect. Therefore the current study was designed to investigate the possible effects of centrally injected CDP-choline on respiratory system and the mediation of the central cholinergic receptors and phospholipase to thromboxane signaling pathway on CDP-choline-induced respiratory effects in anaesthetized rats. Intracerebroventricularly (i.c.v.) administration of CDP-choline induced dose- and time-dependent increased respiratory rates, tidal volume and minute ventilation of male anaesthetized Spraque Dawley rats. İ.c.v. pretreatment with atropine failed to alter the hyperventilation responses to CDP-choline whereas mecamylamine, cholinergic nicotinic receptor antagonist, mepacrine, phospholipase A2 inhibitor, and neomycin phospholipase C inhibitor, blocked completely the hyperventilation induced by CDP-choline. In addition, central pretreatment with furegrelate, thromboxane A2 synthesis inhibitor, also partially blocked CDP-choline-evoked hyperventilation effects. These data show that centrally administered CDP-choline induces hyperventilation which is mediated by activation of central nicotinic receptors and phospholipase to thromboxane signaling pathway.
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http://dx.doi.org/10.1016/j.resp.2014.02.005DOI Listing
May 2014

Protective effect of L-ascorbic acid on nickel induced pulmonary nitrosative stress in male albino rats.

Biometals 2013 Apr 6;26(2):329-36. Epub 2013 Mar 6.

Environmental Health Research Unit, Department of Physiology, Al Ameen Medical College, Bijapur, 586108, Karnataka, India.

Nickel sulfate stimulates inducible nitric oxide synthase (i-NOS) and increases serum nitric oxide concentration by overproduction of reactive nitrogen species due to nitrosative stress. The present study was undertaken to assess possible protective role of L-ascorbic acid as an antioxidant against nickel induced pulmonary nitrosative stress in male albino rats. We studied the effect of the simultaneous treatment with L-ascorbic acid (50 mg/100 g b. wt.; orally) and nickel sulfate (2.0 mg/100 g b. wt.; i.p.) on nitric oxide synthesis by quantitative evaluation of serum i-NOS activities, serum and lung nitric oxide, L-ascorbic acid and protein concentrations of Wistar strain male albino rats. We have further studied histopathological changes in lung tissue after nickel sulfate treatment along with simultaneous exposure of L-ascorbic acid. Nickel sulfate treatment significantly increased the serum i-NOS activity, serum and pulmonary nitric oxide concentration and decreased body weight, pulmonary somatic index, serum and lung L-ascorbic acid and protein concentration as compared to their respective controls. Histopathological changes induced by nickel sulfate showed loss of normal alveolar architecture, inflammation of bronchioles, infiltration of inflammatory cells and patchy congestion of alveolar blood vessels. The simultaneous administration of L-ascorbic acid and nickel sulfate significantly improved all the above biochemical parameters along with histopathology of lung tissues of rats receiving nickel sulfate alone. The study clearly showed a protective role of L-ascorbic acid against nickel induced nitrosative stress in lung tissues.
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http://dx.doi.org/10.1007/s10534-013-9617-3DOI Listing
April 2013

Changes in glutamate transporter expression in mouse forebrain areas following focal ischemia.

Brain Res 2011 Oct 19;1418:93-103. Epub 2011 Aug 19.

Division of Cardiovascular and Neuronal Remodelling, Leeds Institute for Genetics, Health and Therapeutics, University of Leeds, Leeds, LS2 9JT, UK.

Dysfunction of glutamate transporters has been proposed to promote neuronal death in modelled cerebral ischemia. However, these studies have produced conflicting results and the changes in glutamate transporter expression have not yet been examined in a mouse focal ischemic stroke model. This study used quantitative real-time reverse-transcription polymerase chain reaction to examine glutamate transporter mRNA expression in the hippocampus, cortex and striatum in a mouse model of focal ischemic stroke induced by middle cerebral artery occlusion (MCAO). Effects on mRNA expression of glial (GLT-1, GLAST) and neuronal (EAAC1) glutamate transporters in these brain areas were assessed by comparing MCAO brains with sham-operated control brains. Changes in transporter proteins were also assessed by immunohistochemistry using specific antibodies to GLT-1 and GLAST. Following focal ischemia, GLT-1 mRNA expression was decreased significantly in the ipsilateral hippocampus and cortex compared to the sham-operated brains (p<0.05). There were no significant differences in GLAST or EAAC1 mRNA expression between MCAO and sham-operated brains. Immunohistochemistry also confirmed a marked reduction in GLT-1 immunoreactivity in the cortex and hippocampus. Down regulation of GLT-1 in these brain areas may impair normal clearance of synaptically-released glutamate and contribute to neural damage following focal ischemic insult.
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http://dx.doi.org/10.1016/j.brainres.2011.08.029DOI Listing
October 2011

L-ascorbic acid and α tocopherol supplementation and antioxidant status in nickel- or lead-exposed rat brain tissue.

J Basic Clin Physiol Pharmacol 2010 ;21(4):325-46

Environmental Health Research Unit, Department of Physiology, Al Ameen Medical College, Bijapur-586108, Karnataka, India.

We evaluated the effect of L-ascorbic acid and alpha-tocopherol supplementation on plasma and whole brain nitric oxide level and antioxidant status in nickel sulfate- or lead acetate- treated male albino rats. Nitric oxide and lipid peroxide levels in whole brain tissue and plasma increased following nickel and lead treatment but significantly returned to near-normal values upon L-ascorbic acid or alpha-tocopherol supplementation. In brain tissue, antioxidant enzymes--superoxide dismutase, glutathione peroxidase, glutathione peroxidase, and catalase--along with the glutathione level decreased significantly after both treatments but significantly improved upon simultaneous supplementation with L-ascorbic acid or alpha-tocopherol. Lead-treated animals showed a greater improvement with alpha-tocopherol, whereas nickel-treated rats showed a greater improvement with L-ascorbic acid. In both groups, combined supplementation with L-ascorbic acid and alpha-tocopherol did not change the percentage improvement in comparison with supplementation with a single vitamin alone.
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http://dx.doi.org/10.1515/jbcpp.2010.21.4.325DOI Listing
March 2011

Co-localisation of markers for glycinergic and GABAergic neurones in rat nucleus of the solitary tract: implications for co-transmission.

J Chem Neuroanat 2010 Oct 29;40(2):160-76. Epub 2010 Apr 29.

Division of Cardiovascular & Neuronal Remodelling, LIGHT Institute, School of Medicine, University of Leeds, Leeds LS2 9JT, United Kingdom.

Immunoreactive structures visualised with antibodies to glycine were prominent in areas of the nucleus of the solitary tract (NTS) surrounding the tractus solitarius, but scarcer in medial and ventral areas of the nucleus. This contrasted with a higher density, more homogenous distribution of structures labelled for gamma-aminobutyric acid (GABA). Immunolabelling of adjacent semi-thin sections nonetheless indicated a close correspondence between cells and puncta labelled by glycine and GABA antisera in certain NTS areas. With post-embedding electron microscopic immunolabelling, synaptic terminals with high, presumed transmitter levels of glycine were discriminated from terminals containing low, metabolic levels by quantitative analysis of gold particle labelling densities. In a random sample of terminals, 28.5% qualified on this basis as glycinergic (compared to 44.4% GABAergic); these glycinergic terminals targeted mainly dendritic structures and contained pleomorphic vesicles and symmetrical synapses. Serial section analysis revealed few terminals (5.2%) immunoreactive for glycine alone, with 82% of glycinergic terminals also containing high levels of GABA immunoreactivity. No evidence for co-localisation of glycine and glutamate was found. Light, confocal and electron microscopic labelling with antibodies to proteins specific for glycine and GABA synthesis, release and uptake confirmed that glycinergic terminals also containing GABA are found predominantly in more lateral areas of NTS, despite glycine receptors and the 'glial' glycine transporter (GLYT1) being expressed throughout all areas of the nucleus. The data suggest that synaptic terminals in certain functionally distinct areas of NTS co-release both inhibitory amino acids, which may account for the previously reported differential inhibitory effects of glycine and GABA on NTS neurones.
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http://dx.doi.org/10.1016/j.jchemneu.2010.04.001DOI Listing
October 2010

Differential distribution of 5-HT 1A and 5-HT 1B-like immunoreactivities in rat central nucleus of the amygdala neurones projecting to the caudal dorsomedial medulla oblongata.

Brain Res 2010 May 11;1330:20-30. Epub 2010 Mar 11.

Division of Cardiovascular and Neuronal Remodelling, Leeds Institute of Genetics, Health and Therapeutics, Faculty of Medicine, University of Leeds, Leeds, LS2 9JT, UK.

Neurones in the central nucleus of the amygdala (CeA) projecting to the caudal dorsomedial medulla oblongata play a key role in the autonomic expression of emotional behaviour. We have earlier shown that these projections from the CeA contain gamma-aminobutyric acid. The CeA receives a dense serotonergic innervation from the raphé nuclei and expresses several serotonin (5-hydroxytryptamine, 5-HT) receptor subtypes, including the 5-HT(1A) and 5-HT(1B) subtypes. However, there have been no reports on the cellular distribution of these receptors in CeA projection neurones. This study was therefore designed to investigate the localisation of 5-HT(1A) and 5-HT(1B) receptors in CeA projection neurones identified by microinjection of a retrograde tracer, cholera toxin B-subunit (CTb) into the caudal dorsomedial medulla, targeting projections to the nucleus of the solitary tract. A large proportion (approximately 60%) of amygdala neurones retrogradely-labelled with CTb expressed 5-HT(1B) receptor-like immunoreactivity, whereas fewer (approximately 30%) expressed 5-HT(1A) receptor-like immunoreactivity. The retrogradely-labelled neurones positive for 5-HT(1B) receptor were present in both lateral and medial parts of the CeA whereas 5-HT(1A) receptor positive neurones were located mainly in the lateral part of the CeA. Since 5-HT plays an important role in controlling emotional behaviour and the 5-HT(1A) and 5-HT(1B) receptors have been shown to have distinct roles in the regulation of anxiety and depression, the differential expression of these receptors in CeA neurones projecting to the caudal dorsomedial medulla suggests that these projection neurones may act differentially in controlling autonomic expression of emotional behaviour.
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http://dx.doi.org/10.1016/j.brainres.2010.03.009DOI Listing
May 2010

Increased GABA B receptor subtype expression in the nucleus of the solitary tract of the spontaneously hypertensive rat.

J Mol Neurosci 2008 Jun 13;35(2):211-24. Epub 2008 Mar 13.

Division of Cardiovascular and Neuronal Remodelling, LIGHT Institute, Faculty of Medicine & Health, University of Leeds, Worsley Building, Leeds, LS2 9JT, UK.

Expression of GABA(B) receptor messenger RNA (mRNA) in the central nervous system was compared between the spontaneously hypertensive (SHR) and normotensive Wistar Kyoto (WKY) rat. Polymerase chain reaction (PCR) revealed all the isoforms except B1e in cortex, hypothalamus, and medulla oblongata. In the nucleus of the solitary tract (NTS) and ventrolateral medulla (VLM), the B1a-c and 1 g isoforms were present as well as B2. Real-time PCR detected significantly higher levels of B1a (p < 0.01) and B2 (p < 0.05) mRNA in the NTS of SHR compared to WKY. A significant increase in B1a expression (p < 0.05) was detected in VLM. Immunolabeling suggested presynaptic and postsynaptic expression of B1a, B1b, and B2 subtypes throughout the NTS, with significant differences in distribution patterns and labeling between subtypes and between SHR and WKY. These findings suggest that GABA(B) receptors expressed by neurones in NTS may be involved in cardiovascular regulation and that changes in GABA(B) mRNA expression levels may contribute to the hypertensive state in SHR.
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http://dx.doi.org/10.1007/s12031-008-9055-9DOI Listing
June 2008

Central nucleus of amygdala projections to rostral ventrolateral medulla neurones activated by decreased blood pressure.

Eur J Neurosci 2005 Apr;21(7):1921-30

BHF Heart Centre & CRISTAL, School of Medicine, Worsley Building, University of Leeds, Leeds LS2 9JT, UK.

The central nucleus of amygdala (CeA) participates in cardiovascular regulation during emotional behaviour but it has not been established whether any of these effects are mediated through its direct connections to blood pressure-regulating neurones in the rostral ventrolateral medulla (RVLM). The RVLM contains barosensitive neurones that maintain resting blood pressure via their projections to sympathetic preganglionic neurones in the thoracic spinal cord. In this study on rats, we used combined anterograde neuronal tracing of CeA projections with confocal and electron microscopic immunohistochemical detection of phenylethanolamine-N-methyltransferase, the adrenaline-synthesizing enzyme present in C1 catecholamine neurones of the RVLM, and Fos, the protein product of the c-fos proto-oncogene. Fos expression in barosensitive neurones was stimulated by an intravenous infusion of the hypotensive agent sodium nitroprusside. Injection of the tracer biotin dextran amine (10-kDa form) into the CeA resulted in anterograde labelling of axons and varicosities throughout the RVLM without retrograde labelling of somata in any brain area. With confocal microscopy, presumptive CeA terminals were found in close apposition to adrenergic (phenylethanolamine-N-methyltransferase-immunoreactive) and non-adrenergic neurones that displayed Fos-immunoreactive nuclei in response to decreased blood pressure. Electron microscopic analysis confirmed that some labelled terminals of CeA axons made synaptic contact with c-fos-activated adrenergic neurones. The results provide evidence that cardiovascular influences elicited from the CeA during stressful events may be mediated, at least in part, via monosynaptic neural projections to barosensitive sympathetic blood pressure-regulating neurones in the RVLM.
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http://dx.doi.org/10.1111/j.1460-9568.2005.04023.xDOI Listing
April 2005

Differential involvement of oriens/pyramidale interneurones in hippocampal network oscillations in vitro.

J Physiol 2005 Jan 14;562(Pt 1):131-47. Epub 2004 Oct 14.

School of Biomedical Sciences, University of Leeds, Leeds LS2 9JT, UK.

Using whole-cell patch-clamp recordings in conjunction with post hoc anatomy we investigated the physiological properties of hippocampal stratum oriens and stratum pyramidale inhibitory interneurones, before and following the induction of pharmacologically evoked gamma frequency network oscillations. Prior to kainate-induced transient epochs of gamma activity, two distinct classes of oriens interneurones, oriens lacunosum-moleculare (O-LM) and trilaminar cells, showed prominent differences in their membrane and firing properties, as well as in the amplitude and kinetics of their excitatory postsynaptic events. In the active network both types of neurone received a phasic barrage of gamma frequency excitatory inputs but, due to their differential functional integration, showed clear differences in their output patterns. While O-LM cells fired intermittently at theta frequency, trilaminar interneurones discharged on every gamma cycle and showed a propensity to fire spike doublets. Two other classes of fast spiking interneurones, perisomatic targeting basket and bistratified cells, in the active network discharged predominantly single action potentials on every gamma cycle. Thus, within a locally excited network, O-LM cells are likely to provide a theta-frequency patterned output to distal dendritic segments, whereas basket and bistratified cells are involved in the generation of locally synchronous gamma band oscillations. The anatomy and output profile of trilaminar cells suggest they are involved in the projection of locally generated gamma rhythms to distal sites. Therefore a division of labour appears to exist whereby different frequencies and spatiotemporal properties of hippocampal rhythms are mediated by different interneurone subtypes.
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http://dx.doi.org/10.1113/jphysiol.2004.073007DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1665476PMC
January 2005

Increased expression of AMPA receptor subunits in the nucleus of the solitary tract in the spontaneously hypertensive rat.

Brain Res Mol Brain Res 2004 Feb;121(1-2):37-49

School of Medicine, University of Leeds, Leeds LS2 9JT, UK.

The expression of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor subunits GluR1-4 in the nucleus of the solitary tract (NTS) of adult Wistar rats was examined by polymerase chain reaction (PCR), and the neuronal localisation of these receptor subunits in the NTS were confirmed by immunohistochemistry using subunit-specific antibodies. Semi-quantitative PCR was used to investigate differences in AMPA receptor subunit expression between spontaneously hypertensive rats (SH) and age-matched normotensive Wistar Kyoto rats (WKY). All four receptor subunits were expressed in both strains, but compared to WKY, total AMPA receptor and the GluR3 mRNA expressions were significantly higher in SH. No differences were detected in cDNA form the cerebral cortex or cerebellum. Immunolabelling for GluRs 1, 2 and 2/3 in the neuropil relative to neuronal somata in the cardioregulatory areas of the NTS appeared to be increased in SH, with an overall increase in the density of GluR2/3 labelling in the medial and commissural NTS of SH. These results indicate a possible role for changes in AMPA receptor subunit expression in NTS neurones, involving an increase in GluR3 associated with development of hypertension in SH.
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http://dx.doi.org/10.1016/j.molbrainres.2003.11.003DOI Listing
February 2004

Ionotropic glutamate receptor subunit immunoreactivity of vagal preganglionic neurones projecting to the rat heart.

Auton Neurosci 2003 May;105(2):105-17

Institute for Cardiovascular Research, School of Medicine, Worsley Building, University of Leeds, LS2 9JT, Leeds, UK.

The ionotropic glutamate receptor subunits expressed by vagal preganglionic neurones in the rat medulla oblongata were examined by using fluorescence immunolabelling combined with retrograde neuronal tracing. The general population of these neurones in the medulla was identified by intraperitoneal injections of Fluorogold and also with choline acetyltransferase antibodies. Cardiac projecting neurones were specifically identified by applying the fluorescent tracer 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine (DiI) to the heart or by injecting cholera toxin B-subunit into the pericardium. Both tracers labelled populations of neurones lying in the dorsal vagal nucleus, intermediate reticular formation and nucleus ambiguus, and when both tracers were applied simultaneously, approximately 50% of cells were dual-labelled. Control experiments established that the labelling was specific for neurones projecting to the heart. Most vagal preganglionic neurones, including those projecting to the heart, irrespective of their location in the medulla, had a similar profile of glutamate receptor immunoreactivity. Labelling of somata for the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic (AMPA) subunit GluR1 was weak or absent, while labelling with antibodies directed to GluR2, a common sequence of GluR2 and GluR3, and GluR4 was moderate or intense. All neurones studied appeared to express the N-methyl-D-aspartate (NMDA) receptor subunit NR1, and while antibodies recognising the NR2A and NR2B splice variants gave strong labelling, immunoreactivity with a NR2B specific antibody was weaker. Weak to moderate labelling was seen in some neurones using antibodies to the kainate receptor subunits KA2 and GluR5-7. These results are consistent with neurophysiological data indicating the presence of AMPA, NMDA and kainate responses in cardiac vagal preganglionic neurones, and suggest that these neurones are similar to other vagal parasympathetic preganglionic neurones in expressing mainly AMPA receptor subunits GluR2-4.
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http://dx.doi.org/10.1016/S1566-0702(03)00047-XDOI Listing
May 2003

Evidence for peptide co-transmission in retrograde- and anterograde-labelled central nucleus of amygdala neurones projecting to NTS.

Auton Neurosci 2002 Jun;98(1-2):28-32

Institute for Cardiovascular Research, School of Medicine, University of Leeds, UK.

Synaptic terminals in the nucleus of the solitary tract (NTS) from axons originating in the central nucleus of the amygdala (CeA) are known to contain gamma-aminobutyric acid (GABA) immunoreactivity. Here, we have investigated whether such projections contain neuropeptides as putative co-transmitters. Somata in the medial and lateral CeA that were retrogradely labelled with cholera toxin B (CTb) injected into the commissural NTS were found to be immunoreactive for GABA, somatostatin (SOM), neurotensin (NT), vasoactive intestinal polypeptide (VIP) and nitric oxide synthase (NOS). Subpopulations of fibres in the NTS that were anterogradely labelled with biotin dextran amine (BDA) injected into the CeA and examined using both fluorescence and electron microscopy appeared to colocalise somatostatin, but not other neuropeptides. Their varicosities were observed in proximity to NTS neurones that were immunoreactive for the somatostatin receptor sst2A subtype, substance P (SP) NK1 receptor, and the GABAA receptor alpha3, beta1 and gamma2 subunits. This morphological evidence is consistent with the possibility of GABA-somatostatin co-transmission at synapses of some of the CeA projection neurones to NTS that might inhibit cardiovascular reflex responses in response to fear or emotion-related stimuli.
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http://dx.doi.org/10.1016/s1566-0702(02)00026-7DOI Listing
June 2002