Retinitis pigmentosa-associated cystoid macular oedema

Hereditary retinal diseases are now the leading cause of blindness certification in the working age population (age 16-64 years) in England and Wales, of which Retinitis Pigmentosa (RP) is the most common disorder. One complication of RP includes cystoid macular oedema (CMO), causing a reduction of central vision. This thesis begins by reviewing retinal anatomy, retinal function and the visual cycle. It provides a description of the most common inherited retinal disorders (IRD) together with an update on structural, functional and molecular assessment of IRD. The possible underlying causes of RP-associated CMO (RPCMO) are explored, including: 1) Breakdown of the blood-retinal barrier, 2) Failure (or dysfunction) of the pumping mechanism of the retinal pigment epithelium (RPE), 3) Muller cell oedema and dysfunction, 4) Anti-retinal antibodies, and 5) Vitreous traction. Current methods for the diagnosis and monitoring of RP-CMO are discussed. A literature review of all treatments attempted to date will be provided, including: oral and topical carbonic anhydrase inhibitors, oral, topical, intravitreal and periocular steroids, topical non-steroidal anti-inflammatory medications, photocoagulation, vitrectomy with internal limiting membrane peel, oral lutein, and intravitreal anti-vascular endothelial growth factor injections. Extensive explanation is provided regarding the clinical trial undertaken entitled ‘The AMOUR Study’, which stands for ‘Aflibercept for Macular Oedema in Underlying Retinitis Pigmentosa’. The purpose, methods, results and conclusion are provided. Extensive explanation is provided regarding a second retrospective study undertaken entitled ‘The CARAMEL Study’, which stands for ‘Carbonic Anhydrase inhibitors for Retinitis Pigmentosa And Macular oEdema in various Layers’. The purpose, methods, results and conclusion are provided. Everything is drawn together in the discussion and concluding remarks section, including a final chapter on future directions for the treatment of RP-CMO

Time-of-day dependence of neurological deficits induced by sodium nitroprusside in young mice

Sodium nitroprusside (SNP) is widely used in pharmacological studies as a potent vasodilator or a nitric oxide donor. SNP-induced ataxic effects were assessed in mice by the Joulou-Couvoisier test. Swiss albino mice of both genders, 2-8 weeks of age, were acclimated at least for 2 weeks to 12 h light (rest span)/12 h dark (activity span). In 2 and 4 week old mice, maxima of ataxia were found following intraperitoneal administration of a dose ranging from 3 to 3.6 mg.kg-1 SNP at ≈ 1 and 13 HALO (Hours After Light Onset). The sublethal toxicity was statistically dosing-time dependent (χ2 test: P < 0.005). No rhythm was validated in neurotoxicity by cosinor analyses. At the 8th week of post-natal development (PND), SNP-induced ataxia was greatest at ≈ 1 HALO (69% in males vs. 49% in females) and lowest at ≈ 13 HALO (21% in males vs. 11% in females) (χ2 test: P < 0.00001). Cosinor analysis also revealed no statistically significant rhythm in mice injected with 3 or 3.3 mg.kg-1. However, a significant circadian (τ = 24 h) rhythm was detected by adjusted cosinor in 3.6 mg.kg-1-treated mice (P < 0.004). In all studied groups, SNP-induced motor impairment (expressed in %) was lower during the dark than the light phase. Furthermore, there was a non-significant gender-related difference in SNP-induced neuronal toxicity with the males more sensitive than females at every studied PND. The ataxic effects were inversely proportional to the lag time from injection and to the age of animals (with P < 0.05 only between 2 and 8 week old mice). These data indicate that both the administration time and age of the animal significantly affect the neurotoxic effects of SNP

Daily Rhythms of Physiological Parameters in the Dromedary Camel Under Natural and Laboratory Conditions

Camels are well adapted to hot arid environments and can contribute significantly to the economy of developing countries in arid regions of the world. Full understanding of the physiology of camels requires understanding of the internal temporal order of the body, as reflected in daily or circadian rhythms. In the current study, we investigated the daily rhythmicity of 20 physiological variables in camels exposed to natural oscillations of ambient temperature in a desert environment and compared the daily temporal courses of the variables. We also studied the rhythm of core body temperature under experimental conditions with constant ambient temperature in the presence and absence of a light-dark cycle. The obtained results indicated that different physiological variables exhibit different degrees of daily rhythmicity and reach their daily peaks at different times of the day, starting with plasma cholesterol, which peaks 24 minutes after midnight, and ending with plasma calcium, which peaks 3 hours before midnight. Furthermore, the rhythm of core body temperature persisted in the absence of environmental rhythmicity, thus confirming its endogenous nature. The observed delay in the acrophase of core body temperature rhythm under constant conditions suggests that the circadian period is longer than 24 hours. Further studies with more refined experimental manipulation of different variables are needed to fully elucidate the causal network of circadian rhythms in dromedary camels

Dosing-Time Dependent Effects of Sodium Nitroprusside on Cerebral, Renal, and Hepatic Catalase Activity in Mice

To investigate the time dependence of sodium nitroprusside- (NPS-) induced oxidative effects, the authors study the variation of the antioxidant enzyme CAT activity in various tissues after the administration of a single 2.5mg/kg dose of SNP or sodium chloride (NaCl 0.9%). For each of the two dosing times (1 and 13 hours after light onset, HALO, which correspond to the beginning of diurnal rest span and of nocturnal activity span of mice, resp.), brain, kidney, and liver tissues were excised from animals at 0, 1, 3, 6, 9, 12, 24, and 36 h following the drug administration and CAT activity was assayed.The results suggest that SNP-induced stimulation of CAT activity is greater in all three tissues when the drug is administered at 1 HALO than at 13 HALO. Two-way ANOVA revealed that CAT activity significantly (P \u3c 0.004) varied as a function of the sampling time but not of the treatment in all three tissues. Moreover, a statistically significant (P \u3c 0.004) interaction between the organ sampling-time and the SNP treatment was revealed in kidney regardless of the dosing time, whereas a highly significant (P \u3c 0.0002) interaction was validated in liver only in animals injected at 13 HALO

Large Formal Wikis: Issues and Solutions

We present several steps towards large formal mathematical wikis. The Coq proof assistant together with the CoRN repository are added to the pool of systems handled by the general wiki system described in \cite{DBLP:conf/aisc/UrbanARG10}. A smart re-verification scheme for the large formal libraries in the wiki is suggested for Mizar/MML and Coq/CoRN, based on recently developed precise tracking of mathematical dependencies. We propose to use features of state-of-the-art filesystems to allow real-time cloning and sandboxing of the entire libraries, allowing also to extend the wiki to a true multi-user collaborative area. A number of related issues are discussed.Comment: To appear in The Conference of Intelligent Computer Mathematics: CICM 201

mTOR signaling: implications for cancer and anticancer therapy

Mounting evidence links deregulated protein synthesis to tumorigenesis via the translation initiation factor complex eIF4F. Components of this complex are often overexpressed in a large number of cancers and promote malignant transformation in experimental systems. mTOR affects the activity of the eIF4F complex by phosphorylating repressors of the eIF4F complex, the eIF4E binding proteins. The immunosuppressant rapamycin specifically inhibits mTOR activity and retards cancer growth. Importantly, mutations in upstream negative regulators of mTOR cause hamartomas, haemangiomas, and cancers that are sensitive to rapamycin treatment. Such mutations lead to increased eIF4F formation and consequently to enhanced translation initiation and cell growth. Thus, inhibition of translation initiation through targeting the mTOR-signalling pathway is emerging as a promising therapeutic option

An approach to analyse the specific impact of rapamycin on mRNA-ribosome association

<p>Abstract</p> <p>Background</p> <p>Recent work, using both cell culture model systems and tumour derived cell lines, suggests that the differential recruitment into polysomes of mRNA populations may be sufficient to initiate and maintain tumour formation. Consequently, a major effort is underway to use high density microarray profiles to establish molecular fingerprints for cells exposed to defined drug regimes. The aim of these pharmacogenomic approaches is to provide new information on how drugs can impact on the translational read-out within a defined cellular background.</p> <p>Methods</p> <p>We describe an approach that permits the analysis of de-novo mRNA-ribosome association in-vivo during short drug exposures. It combines hypertonic shock, polysome fractionation and high-throughput analysis to provide a molecular phenotype of translationally responsive transcripts. Compared to previous translational profiling studies, the procedure offers increased specificity due to the elimination of the drugs secondary effects (e.g. on the transcriptional read-out). For this pilot "proof-of-principle" assay we selected the drug rapamycin because of its extensively studied impact on translation initiation.</p> <p>Results</p> <p>High throughput analysis on both the light and heavy polysomal fractions has identified mRNAs whose re-recruitment onto free ribosomes responded to short exposure to the drug rapamycin. The results of the microarray have been confirmed using real-time RT-PCR. The selective down-regulation of TOP transcripts is also consistent with previous translational profiling studies using this drug.</p> <p>Conclusion</p> <p>The technical advance outlined in this manuscript offers the possibility of new insights into mRNA features that impact on translation initiation and provides a molecular fingerprint for transcript-ribosome association in any cell type and in the presence of a range of drugs of interest. Such molecular phenotypes defined pre-clinically may ultimately impact on the evaluation of a particular drug in a living cell.</p

Downsizing a human inflammatory protein to a small molecule with equal potency and functionality

A significant challenge in chemistry is to rationally reproduce the functional potency of a protein in a small molecule, which is cheaper to manufacture, non-immunogenic, and also both stable and bioavailable. Synthetic peptides corresponding to small bioactive protein surfaces do not form stable structures in water and do not exhibit the functional potencies of proteins. Here we describe a novel approach to growing small molecules with protein-like potencies from a functionally important amino acid of a protein. A 77-residue human inflammatory protein (complement C3a) important in innate immunity is rationally transformed to equipotent small molecules, using peptide surrogates that incorporate a turn-inducing heterocycle with correctly positioned hydrogen-bond-accepting atoms. Small molecule agonists (molecular weigh