• Na+/Ca2+ exchange current INa/Ca) and sarcoplasmic reticulum (SR) Ca2+ release in catecholamine-induced cardiac hypertrophy.

      Hussain, Munir; Chorvatova, A.; Hart, G. (2004)
      Catecholamines that accompany acute physiological stress are also involved in mediating the development of hypertrophy and failure. However, the cellular mechanisms involved in catecholamine-induced cardiac hypertrophy, particularly Ca2+ handling, are largely unknown. We therefore investigated the effects of cardiac hypertrophy, produced by isoprenaline, on INa/Ca and sarcoplasmic reticulum (SR) function in isolated myocytes. Methods: INa/Ca was studied in myocytes from Wistar rats, using descending (+80 to ¿110 mV) voltage ramps under steady state conditions. Myocytes were also loaded with fura-2 and either field stimulated or voltage clamped to assess [Ca2+]i and SR Ca2+ content. Results: Ca2+-dependent, steady state INa/Ca density was increased in hypertrophied myocytes (P<0.05). Ca2+ release from the SR was also increased, whereas resting [Ca2+]i and the rate of decline of [Ca2+]i to control levels were unchanged. SR Ca2+ content, estimated by using 10.0 mmol/l caffeine, was also significantly increased in hypertrophied myocytes, but only when myocytes were held and stimulated from their normal resting potential (¿80 mV) but not from ¿40 mV. However, the rate of decline of caffeine-induced Ca2+ transients or INa/Ca was not significantly different between control and hypertrophied myocytes. Ca2+-dependence of INa/Ca, examined by comparing the slope of the descending phase of the hysteresis plots of INa/Ca vs. [Ca2+]i, was also similar in the two groups of cells. Conclusion: Data show that SR Ca2+ release and SR Ca2+ content were increased in hypertrophied myocytes, despite an increase in the steady state INa/Ca density. The observation that increased SR function occurred only when myocytes were stimulated from ¿80 mV suggests that Na+ influx may play a role in altering Ca2+ homeostasis in hypertrophied cardiac muscle, possibly through increased reverse Na+/Ca2+ exchange, particularly at low stimulation frequencies.
    • Nano-encapsulation of a novel anti-Ran-GTPase peptide for blockade of regulator of chromosome condensation 1 (RCC1) function in MDA-MB-231 breast cancer cells

      Haggag, Y.A.; Matchett, K.B.; Dakir, El-Habib; Buchanan, P.; Osman, M.A.; Elgizawy, S.A.; El-Tanani, Mohamed; Faheem, A.M.; McCarron, P.A. (2017-04)
      Ran is a small ras-related GTPase and is highly expressed in aggressive breast carcinoma. Overexpression induces malignant transformation and drives metastatic growth. We have designed a novel series of anti-Ran-GTPase peptides, which prevents Ran hydrolysis and activation, and although they display effectiveness in silico, peptide activity is suboptimal in vitro due to reduced bioavailability and poor delivery. To overcome this drawback, we delivered an anti-Ran-GTPase peptide using encapsulation in PLGA-based nanoparticles (NP). Formulation variables within a double emulsion solvent evaporation technique were controlled to optimise physicochemical properties. NP were spherical and negatively charged with a mean diameter of 182–277 nm. Peptide integrity and stability were maintained after encapsulation and release kinetics followed a sustained profile. We were interested in the relationship between cellular uptake and poly(ethylene glycol) (PEG) in the NP matrix, with results showing enhanced in vitro uptake with increasing PEG content. Peptide-loaded, pegylated (10% PEG)-PLGA NP induced significant cytotoxic and apoptotic effects in MDA-MB-231 breast cancer cells, with no evidence of similar effects in cells pulsed with free peptide. Western blot analysis showed that encapsulated peptide interfered with the proposed signal transduction pathway of the Ran gene. Our novel blockade peptide prevented Ran activation by blockage of regulator of chromosome condensation 1 (RCC1) following peptide release directly in the cytoplasm once endocytosis of the peptide-loaded nanoparticle has occurred. RCC1 blockage was effective only when a nanoparticulate delivery approach was adopted.
    • Nano-Scale Observations of Tattoo Pigments in Skin by Atomic Force Microscopy

      Grant, Colin A.; Twigg, Peter C.; Tobin, Desmond J. (2015-03-26)
      In this study, we have shown how particles in carbon black tattoo ink accumulate in the human skin dermis using fine-resolution atomic force microscopy, with which a single ink particle in the collagenous network can be imaged. This information further demonstrates that tattoo inks are nano-particles. Further, we have deposited a commercially available tattoo ink on a glass slide and calculated a range of volumes for single ink particles.
    • Nanoparticle labels for pathogen detection through nucleic acid amplification tests

      Drake, Philip; Chen, Y-C.; Lehmann, I.; Jiang, P-S. (2015-08)
      Magnetic nanoparticles and surface-enhanced Raman scattering (SERS) active nanoparticles were coated with short chain DNA tags. These were then used to identify a target bacterial DNA sequence. The tags function as primers in a standard PCR with the reverse primers and forward primers on the SERS nanoparticles and magnetic nanoparticles, respectively. During the PCR cycles, a composite nanostructure is formed that is both magnetically responsive and SERS active. After magnetic trapping, the intensity of the SERS signal can be related back to the concentration of the target DNA. A test assay was performed that showed a detection limit (based on the signal to noise ratio) of less than 3 zeptomole (41 pg/L). For comparison, a PCR assay based on the standard SYBR Green method was performed. This used the same primers and target DNA and had a detection limit of 10 attomoles (138 ng/L), 3,000 times less sensitive. The work documents the proof of principle study and shows for the first time the use of SERS-NP labels in the quantification of nucleic acid amplification tests and PCR.
    • Nanoparticles for post-infarct ventricular remodeling

      Dong, C.; Ma, A.; Shang, Lijun (2018-12)
      In recent years, tremendous progress has been made in the treatment of acute myocardial infarction (AMI), but pathological ventricular remodeling often causes survivors to suffer from fatal heart failure. Currently, there is no effective therapy to attenuate ventricular remodeling. Recently, nanoparticles-based drug delivery system is widely applied in biomedicine especially in cancer and liver fibrosis, owing to its excellent physical, chemical, and biological properties. Therefore, using nanoparticles as delivery vehicles of small molecules, polypeptides, etc to improve post-infarct ventricular remodeling are expected. In this review, we summarized the updated researches in this fast-growing area and suggested further works needed.
    • Nanoparticles in Biomedicine and Medicine, and Possible Clinical Toxicological Application of Peripheral Lymphocytes in the Risk Assessment Process for Susceptible Disease State Individuals

      Najafzadeh, Mojgan; Anderson, Diana (2017-11-13)
      Nanoparticle usage has emerged in the medical field as a technology well-suited to the diagnosis and treatment of a variety of disease states. The distinctive characteristics of engineered nanoparticles (ENPs) such as higher surface-area-to-volume ratios find various applications in personal care products, food packaging, drug-delivery systems, therapeutics, biosensors and others. The exponential increase in ENP-containing consumer products in the last decade has also increased their inadvertent release into the environment and the debate relating to their adverse effects on human and environmental health. The use of NPs for different functions in human studies has significantly increased the application of NPs in biomedicine, for instance, imaging of cell and tissues, drug delivery and sensing of target molecules. These nanomaterials have been investigated for the treatment and detection of various pathological conditions. There are suitable biological systems now available in man using peripheral blood lymphocytes to determine the effect of NPs in various disease states.
    • Nanoparticles of chitosan conjugated to organo-ruthenium complexes

      Wang, Y.; Pitto-Barry, Anaïs; Habtemariam, A.; Romero-Canelón, I.; Sadler, P.J.; Barry, Nicolas P.E. (2016)
      The synthesis of nanoparticles of conjugates of caffeic acid-modified chitosan with ruthenium arene complexes is described. The chemical structure and physical properties of the nanoparticles were characterised by electronic absorption spectroscopy (UV-vis), Fourier transform infrared spectroscopy (FT-IR), 1H NMR spectroscopy, dynamic light scattering (DLS), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), and circular dichroism (CD) analysis. The multi-spectral results revealed that caffeic acid is covalently bound to chitosan and chelates to {Ru(p-cymene)Cl}+. The DLS studies indicated that the Ru–caffeic acid modified chitosan nanoparticles are well-defined and of nanometre size. Such well-defined nanocomposites of chitosan and metal complexes might find a range of applications, for example in drug delivery.
    • Nanosizing of hydrocortisone using microfluidic reactors.

      Ali, H.R.H.; York, Peter; Blagden, Nicholas (2008)
      The formulation of poorly water-soluble drugs is a challenging problem within pharmaceutical development. Recently, formulation using nanoparticles was highlighted as showing great potential to improve the dissolution and solubility characteristics of poorly water soluble drugs.
    • A natural solution to photoprotection and isolation of the potent polyene antibiotic, marinomycin A

      Bailey, C.S.; Zarins-Tutt, J.S.; Agbo, M.; Gao, H.; Diego-Taboada, A.; Gan, M.; Hamed, Refaat B.; Abraham, E.R.; Mckenzie, G.; Evans, P.A.; et al. (2019-08-28)
      The photoprotection and isolation of marinomycin A using sporopollenin exine capsules (SpECs) derived from the spores of the plant Lycopodium clavatum is described. The marinomycins have a particularly short half-life in natural light, which severely impacts their potential biological utility given that they display potent antibiotic and anticancer activity. The SpEC encapsulation of the marinomycin A dramatically increases the half-life of the polyene macrodiolide to the direct exposure to UV radiation by several orders of magnitude, thereby making this a potentially useful strategy for other light sensitive bioactive agents. In addition, we report that the SpECs can also be used to selectively extract culture broths that contain the marinomycins, which provides a significantly higher recovery than with conventional XAD resins and provides concomitant photoprotection.
    • Nature versus design: the conformational propensities of D-amino acids and the importance of side chain chirality

      Towse, Clare-Louise; Hopping, G.G.; Vulovic, I.M.; Daggett, V. (2014-11-27)
      D-amino acids are useful building blocks for de novo peptide design and they play a role in aging-related diseases associated with gradual protein racemization. For amino acids with achiral side chains, one should be able to presume that the conformational propensities of L- and D-amino acids are a reflection of one another due to the straightforward geometric inversion at the Cα atom. However, this presumption does not account for the directionality of the backbone dipole and the inverted propensities have never been definitively confirmed in this context. Furthermore, there is little known of how alternative side chain chirality affects the backbone conformations of isoleucine and threonine. Using a GGXGG host-guest pentapeptide system, we have completed exhaustive sampling of the conformational propensities of the D-amino acids, including D-allo-isoleucine and D-allo-threonine, using atomistic molecular dynamics simulations. Comparison of these simulations with the same systems hosting the cognate L-amino acids verifies that the intrinsic backbone conformational propensities of the D-amino acids are the inverse of their cognate L-enantiomers. Where amino acids have a chiral center in their side chain (Thr, Ile) the β-configuration affects the backbone sampling, which in turn can confer different biological properties.
    • Near infra red spectroscopy as a multivariate process analytical tool for predicting pharmaceutical co-crystal concentration

      Wood, Clive; Alwati, Abdolati; Halsey, S.A.; Gough, Timothy D.; Brown, Elaine C.; Kelly, Adrian L.; Paradkar, Anant R. (2016-09-10)
      The use of near infra red spectroscopy to predict the concentration of two pharmaceutical co-crystals; 1:1 ibuprofen – nicotinamide (IBU-NIC) and 1:1 carbamazepine – nicotinamide (CBZ-NIC) has been evaluated. A Partial Least Squares (PLS) regression model was developed for both co-crystal pairs using sets of standard samples to create calibration and validation data sets with which to build and validate the models. Parameters such as the root mean square error of calibration (RMSEC), root mean square error of prediction (RMSEP) and correlation coefficient were used to assess the accuracy and linearity of the models. Accurate PLS regression models were created for both co-crystal pairs which can be used to predict the co-crystal concentration in a powder mixture of the co-crystal and the active pharmaceutical ingredient (API). The IBU-NIC model had smaller errors than the CBZ-NIC model, possibly due to the complex CBZ-NIC spectra which could reflect the different arrangement of hydrogen bonding associated with the co-crystal compared to the IBU-NIC co-crystal. These results suggest that NIR spectroscopy can be used as a PAT tool during a variety of pharmaceutical co-crystal manufacturing methods and the presented data will facilitate future offline and in-line NIR studies involving pharmaceutical co-crystals.
    • A needs assessment for a minor eye condition service within Leeds, Bradford and Airedale, UK

      Swystun, Alexander G.; Davey, Christopher J. (2019-08)
      Background: There are a number of limitations to the present primary eye care system in the UK. Patients with minor eye conditions typically either have to present to their local hospital or GP, or face a charge when visiting eye care professionals (optometrists). Some areas of the UK have commissioned enhanced community services to alleviate this problem; however, many areas have not. The present study is a needs assessment of three areas (Leeds, Airedale and Bradford) without a Minor Eye Conditions Service (MECS), with the aim of determining whether such a service is clinically or economically viable. Method: A pro forma was developed for optometrists and practice staff to complete when a patient presented whose reason for attending was due to symptoms indicative of a problem that could not be optically corrected. This form captured the reason for visit, whether the patient was seen, the consultation funding, the outcome and where the patient would have presented to if the optometrists could not have seen them. Optometrists were invited to participate via Local Optical Committees. Results were submitted via a Google form or a Microsoft Excel document and were analysed in Microsoft Excel. Results: Seventy-five percent of patients were managed in optometric practice. Nine and 16% of patients required subsequent referral to their General Practitioner or hospital ophthalmology department, respectively. Should they not have been seen, 34% of patients would have presented to accident and emergency departments and 59% to their general practitioner. 53% of patients paid privately for the optometrist appointment, 28% of patients received a free examination either through use of General Ophthalmic Service sight tests (9%) or optometrist good will (19%) and 19% of patients did not receive a consultation and were redirected to other providers (e.g. pharmacy, accident and emergency or General Practitioner). 88% of patients were satisfied with the level of service. Cost-analyses revealed a theoretical cost saving of £3198 to the NHS across our sample for the study period, indicating cost effectiveness. Conclusions: This assessment demonstrates that a minor eye condition service in the local areas would be economically and clinically viable and well received by patients.
    • Negative feedback regulation of the ERK1/2 MAPK pathway

      Lake, D.; Corrêa, Sonia A.L.; Muller, Jurgen (2016)
      The extracellular signal-regulated kinase 1/2 (ERK1/2) mitogen-activated protein kinase (MAPK) signalling pathway regulates many cellular functions, including proliferation, differentiation, and transformation. To reliably convert external stimuli into specific cellular responses and to adapt to environmental circumstances, the pathway must be integrated into the overall signalling activity of the cell. Multiple mechanisms have evolved to perform this role. In this review, we will focus on negative feedback mechanisms and examine how they shape ERK1/ 2 MAPK signalling. We will first discuss the extensive number of negative feedback loops targeting the different components of the ERK1/2 MAPK cascade, specifically the direct posttranslational modification of pathway components by downstream protein kinases and the induction of de novo gene synthesis of specific pathway inhibitors. We will then evaluate how negative feedback modulates the spatiotemporal signalling dynamics of the ERK1/2 pathway regarding signalling amplitude and duration as well as subcellular localisation. Aberrant ERK1/2 activation results in deregulated proliferation and malignant transformation in model systems and is commonly observed in human tumours. Inhibition of the ERK1/2 pathway thus represents an attractive target for the treatment of malignant tumours with increased ERK1/2 activity. We will, therefore, discuss the effect of ERK1/2 MAPK feedback regulation on cancer treatment and how it contributes to reduced clinical efficacy of therapeutic agents and the development of drug resistance.
    • Neither here nor there: localizing conflicting visual attributes

      Whitaker, David J.; Badcock, D.R.; McGraw, Paul V.; Skillen, Jennifer (2003)
      Natural visual scenes are a rich source of information. Objects often carry luminance, colour, motion, depth and textural cues, each of which can serve to aid detection and localization of the object within a scene. Contemporary neuroscience presumes a modular approach to visual analysis in which each of these attributes are processed within ostensibly independent visual streams and are transmitted to geographically distinct and functionally dedicated centres in visual cortex (van Essen & Maunsell, 1983; Zihl, von Cramon & Mai, 1983; Maunsell & Newsome, 1987; Tootell, Hadjikhani, Mendola, Marrett & Dale, 1998). In the present study we ask how the visual system localizes objects within this framework. Specifically, we investigate how the visual system assigns a unitary location to objects defined by multiple stimulus attributes, where such attributes provide conflicting positional cues. The results show that conflicting sources of visual information can be effortlessly combined to form a global estimate of spatial position, yet, this conflation of visual attributes is achieved at a cost to localization accuracy. Furthermore, our results suggest that the visual system assigns more perceptual weight (Landy, 1993; Landy & Kojima, 2001) to visual attributes which are reliably related to object contours.
    • Neolithic zoomorphic vessels from Eastern Macedonia, Greece: Issues of function

      Marangou, C.; Stern, Ben (2009)
      Five fragments of Late Neolithic clay zoomorphic vessels from northern Greece have been analysed for organic residues by gas chromatography - mass spectrometry. The results showed that the containers had been used in connection with a number of substances, in particular lower terpenoids, an oil or fat, possibly fossil fuel and in one case possibly beeswax. The paper considers likely interpretations of such combinations of materials in relation to possible functions of these symbolically enhanced artefacts. It appears that substances may have been used in the vessels because of their aromatic and/or medicinal and combustible properties, possibly in order to produce light, fragrance and/or smoke.
    • Nerve Growth Factor Partially Recovers Inflamed Skin from Stress-Induced Worsening in Allergic Inflammation.

      Peters, E.M.J.; Liezman, C.; Spatz, K.; Daniltchenko, M.; Ricardo, J.; Gimenez-Rivera, A.; Hendrix, S.; Botchkarev, Vladimir A.; Brandner, J.M.; Klapp, B.F. (2011)
      Neuroimmune dysregulation characterizes atopic disease, but its nature and clinical impact remain ill-defined. Induced by stress, the neurotrophin nerve growth factor (NGF) may worsen cutaneous inflammation. We therefore studied the role of NGF in the cutaneous stress response in a mouse model for atopic dermatitis–like allergic dermatitis (AlD). Combining several methods, we found that stress increased cutaneous but not serum or hypothalamic NGF in telogen mice. Microarray analysis showed increased mRNAs of inflammatory and growth factors associated with NGF in the skin. In stress-worsened AlD, NGF-neutralizing antibodies markedly reduced epidermal thickening together with NGF, neurotrophin receptor (tyrosine kinase A and p75 neurotrophin receptor), and transforming growth factor-β expression by keratinocytes but did not alter transepidermal water loss. Moreover, NGF expression by mast cells was reduced; this corresponded to reduced cutaneous tumor necrosis factor-α (TNF-α) mRNA levels but not to changes in mast cell degranulation or in the T helper type 1 (Th1)/Th2 cytokine balance. Also, eosinophils expressed TNF receptor type 2, and we observed reduced eosinophil infiltration after treatment with NGF-neutralizing antibodies. We thus conclude that NGF acts as a local stress mediator in perceived stress and allergy and that increased NGF message contributes to worsening of cutaneous inflammation mainly by enhancing epidermal hyperplasia, pro-allergic cytokine induction, and allergy-characteristic cellular infiltration.
    • Nerve guides manufactured from photocurable polymers to aid peripheral nerve repair

      Pateman, C.J.; Harding, A.J.; Glen, A.; Taylor, C.S.; Christmas, C.R.; Robinson, P.P.; Rimmer, Stephen; Boissonade, F.M.; Claeyssens, F.; Haycock, J.W. (2015)
      The peripheral nervous system has a limited innate capacity for self-repair following injury, and surgical intervention is often required. For injuries greater than a few millimeters autografting is standard practice although it is associated with donor site morbidity and is limited in its availability. Because of this, nerve guidance conduits (NGCs) can be viewed as an advantageous alternative, but currently have limited efficacy for short and large injury gaps in comparison to autograft. Current commercially available NGC designs rely on existing regulatory approved materials and traditional production methods, limiting improvement of their design. The aim of this study was to establish a novel method for NGC manufacture using a custom built laser-based microstereolithography (muSL) setup that incorporated a 405 nm laser source to produce 3D constructs with approximately 50 mum resolution from a photocurable poly(ethylene glycol) resin. These were evaluated by SEM, in vitro neuronal, Schwann and dorsal root ganglion culture and in vivo using a thy-1-YFP-H mouse common fibular nerve injury model. NGCs with dimensions of 1 mm internal diameter x 5 mm length with a wall thickness of 250 mum were fabricated and capable of supporting re-innervation across a 3 mm injury gap after 21 days, with results close to that of an autograft control. The study provides a technology platform for the rapid microfabrication of biocompatible materials, a novel method for in vivo evaluation, and a benchmark for future development in more advanced NGC designs, biodegradable and larger device sizes, and longer-term implantation studies.
    • Neural network based modelling and control of batch reactor.

      Mujtaba, Iqbal M.; Aziz, Norashid; Hussain, M.A. (2006)
      The use of neural networks (NNs) in all aspects of process engineering activities, such as modelling, design, optimization and control has considerably increased in recent years (Mujtaba and Hussain, 2001). In this work, three different types of nonlinear control strategies are developed and implemented in batch reactors using NN techniques. These are generic model control (GMC), direct inverse model control (DIC) and internal model control (IMC) strategies. Within the control strategies, NNs have been used as dynamic estimator, dynamic model (forward model) and control (inverse model). An exothermic complex reaction scheme in a batch reactor is considered to explain all these control strategies and their robustness. A dynamic optimization problem with a simple model is solved a priori to obtain optimal operation policy in terms of the reactor temperature with an objective to maximize the desired product in a given batch time. The resulting optimal temperature policy is used as set-point in the control study. All types of controllers performed well in tracking the optimal temperature profile and achieving target conversion to the desired product. However, the NNs used in DIC and IMC controllers need training beyond the nominal operating condition to cope with uncertainties better.
    • Neural Wiskott-Aldrich syndrome protein modulates Wnt signaling and is required for hair follicle cycling in mice

      Lyubimova, A.; Garber, J.J.; Upadhyay, G.; Sharov, A.A.; Anastasoaie, F.; Yajnik, V.; Cotsarelis, G.; Dotto, G.P.; Botchkarev, Vladimir A.; Snapper, S.B. (2010)
      The Rho family GTPases Cdc42 and Rac1 are critical regulators of the actin cytoskeleton and are essential for skin and hair function. Wiskott-Aldrich syndrome family proteins act downstream of these GTPases, controlling actin assembly and cytoskeletal reorganization, but their role in epithelial cells has not been characterized in vivo. Here, we used a conditional knockout approach to assess the role of neural Wiskott-Aldrich syndrome protein (N-WASP), the ubiquitously expressed Wiskott-Aldrich syndrome-like (WASL) protein, in mouse skin. We found that N-WASP deficiency in mouse skin led to severe alopecia, epidermal hyperproliferation, and ulceration, without obvious effects on epidermal differentiation and wound healing. Further analysis revealed that the observed alopecia was likely the result of a progressive and ultimately nearly complete block in hair follicle (HF) cycling by 5 months of age. N-WASP deficiency also led to abnormal proliferation of skin progenitor cells, resulting in their depletion over time. Furthermore, N-WASP deficiency in vitro and in vivo correlated with decreased GSK-3beta phosphorylation, decreased nuclear localization of beta-catenin in follicular keratinocytes, and decreased Wnt-dependent transcription. Our results indicate a critical role for N-WASP in skin function and HF cycling and identify a link between N-WASP and Wnt signaling. We therefore propose that N-WASP acts as a positive regulator of beta-catenin-dependent transcription, modulating differentiation of HF progenitor cells.
    • Neurodegenerative Diseases: Pathology, Mechanisms and Potential Therapeutic Targets

      Beart, P.; Robinson, M.; Rattray, Marcus; Maragakis, N. (2017-08)
      Provides a timely overview of critical advances in molecular and cellular neurobiology, covers key methodologies driving progress, and highlights key future directions for research on neuronal injury and neurodegeneration relevant to neuronal brain pathologies. The editors bring together contributions from internationally recognized workers in the field to provide an up to date account of how and why molecular and cellular neurobiology is such an important area for clinical neuroscience. Understanding the molecular aspects of a number of neurodegenerative conditions such as Parkinson's or Alzheimer's disease for the purpose of improving patient management remains a major challenge of neurobiology be it from the basic or clinical perspective. A strategic evaluation of research contributions and the power of modern methods will help advance knowledge over the next years.