Altered visual processing in a rodent model of Attention-Deficit Hyperactivity Disorder.

A central component of Attention-Deficit Hyperactivity Disorder (ADHD) is increased distractibility, which is linked to the superior colliculus (SC) in a range of species, including humans. Furthermore, there is now mounting evidence of altered collicular functioning in ADHD and it is proposed that a hyper-responsive SC could mediate the main symptoms of ADHD, including distractibility. In the present study we have provided a systematic characterization of the SC in the most commonly used and well-validated animal model of ADHD, the spontaneously hypertensive rat (SHR). We examined collicular-dependent orienting behavior, local field potential (LFP) and multiunit responses to visual stimuli in the anesthetized rat and morphological measures in the SHR in comparison to the Wistar Kyoto (WKY) and Wistar (WIS). We found that SHRs remain responsive to a repeated visual stimulus for more presentations than control strains and have a longer response duration. In addition, LFP and multiunit activity within the visually responsive superficial layers of the SC showed the SHR to have a hyper-responsive SC relative to control strains, which could not be explained by altered functioning of the retinocollicular pathway. Finally, examination of collicular volume, neuron and glia densities and glia:neuron ratio revealed that the SHR had a reduced ratio relative to the WKY which could explain the increased responsiveness. In conclusion, this study demonstrates strain-specific changes in the functioning and structure of the SC in the SHR, providing convergent evidence that the SC might be dysfunctional in ADHD.

Preparation and evaluation of microspheres prepared from novel polyester-ibuprofen conjugates blended with non-conjugated ibuprofen.

A novel polyester, poly(glycerol-adipate-co-omega-pentadecalactone) (PGA-co-PL), was conjugated with a model drug, ibuprofen, through the free hydroxyl groups of the former and the free carboxyl group of the latter at various levels of substitution. The conjugated material was processed into microspheres by both emulsion solvent evaporation and spray-drying methods. Samples of conjugated material were also blended with non-conjugated drug and the microspheres produced were evaluated by various methods. Morphologically, the microspheres produced were satisfactory. However, there was some initial burst drug release from all samples, probably due to the presence of non-conjugated drug. Subsequent drug release was very slow due to the relative stability of the covalent bonding of the drug-polyester conjugate. Stability tests showed that storage at high relative humidity resulted in increased burst release.

Synthesis and evaluation of novel polyester-ibuprofen conjugates for modified drug release.

Ibuprofen was conjugated at different levels to a novel polyester, poly(glycerol-adipate-co-omega-pentadecalactone) (PGA-co-PL), via an ester linkage to form a prodrug. The conjugates were characterized by differential scanning calorimetry (DSC), nuclear magnetic resonance (NMR), infrared (IR), gel permeation chromatography (GPC), ultraviolet (UV), and high-performance liquid chromatography (HPLC). The conjugates had a molecular weight between 18 and 24 kDa, and there was a suppression of the free hydroxyl groups within the conjugated polymer. DSC scans showed a lowering of the melting point (T(m)) when compared with the polyester alone and a difference in the number and area of T(m) peaks. Drug release studies showed an initial burst release (13-18%) followed thereafter by very slow release (maximum 35% after 18 days). Continuous work may produce ester-linked conjugates that are sufficiently labile to allow for complete release of ibuprofen over the time period studied.

Cytotoxicity evaluation of anionic nanoliposomes and nanolipoplexes prepared by the heating method without employing volatile solvents and detergents.

Submicron lipid vesicles (nanoliposomes) are being used as carriers of bioactive compounds. In addition, complexes of nanoliposomes and nucleic acids (nanolipoplexes) are promising tools for the treatment of cancer, and viral and genetic disorders. Toxicity of some of these formulations, however, still remains a concern in their clinical utilisation. To address this problem, anionic liposomes were prepared by two different techniques, the conventional thin-film method, and the heating method (HM), in which no volatile organic solvent or detergent is used. An anionic nanolipoplex was constructed by incorporating plasmid DNA (pcDNA3.1/His B/lacZ) into the HM-nanoliposomes by the mediation of calcium. The toxicity of the nanoliposomes, with and without plasmid and Ca2+, was assessed using a human bronchial epithelial cell line (16HBE14o-) in the presence of serum. Cytotoxicity evaluations performed by two different assays (i.e. NRU and MTT) indicated that HM-nanoliposomes were completely non-toxic in the cell-line tested, whereas conventional liposomes revealed significant levels of toxicity. This may be due to the presence of trace amounts of chloroform and/or methanol applied during their preparation. Similar results were obtained for different sizes of lipid vesicles (prepared by 100 nm and 400 nm pore-size filters). In addition, it was observed that incorporation of DNA (15 microg/ 285 microg lipid) and Ca2+ (50 mM) to the nanoliposomes did not have any effect on their cytotoxicities. These findings indicate that the HM-liposomes have great potential as non-toxic delivery vehicles in human gene therapy and drug delivery applications while liposomes made using organic solvents should be used with caution.

Evaluation of ibuprofen-loaded microspheres prepared from novel copolyesters.

The utility of two novel linear random copolyesters to encapsulate and control the release of ibuprofen, via microspheres, was investigated. Various manufacturing parameters, including temperature, disperse phase volume and polymer:ibuprofen ratios were altered during the microsphere production. The effects of these changes on the morphological characteristics of the microspheres, yield, drug loading, encapsulation efficiency and drug release rates were examined. The diameter of the microspheres ranged from 36 to 89 microm and showed both smooth and ridged surfaces. Microsphere diameter was probably determined by the internal phase volume, while surface morphology was controlled by manufacturing temperature. Greater encapsulation efficiency was obtained by increasing the polymer:ibuprofen ratio and by reducing the internal phase volume. For all batches there was an initial burst drug release into phosphate buffer (pH 7.4) over the first 2-4h, which was followed by a much slower release rate over the remaining time period. Drug release rates during both these phases were dependent upon the amount and nature of the polymer in the microspheres, noting that the more hydrophilic polymer provided faster release rates. Ibuprofen solubility appeared to play a dominant role in controlling release, although both encapsulation efficiency and microsphere morphology were also contributing factors.

A review of scanning probe microscopy investigations of liposome-DNA complexes.

Liposome-DNA complexes are one of the most promising systems for the protection and delivery of nucleic acids to combat neoplastic, viral, and genetic diseases. In addition, they are being used as models in the elucidation of many biological phenomena such as viral infection and transduction. In order to understand these phenomena and to realize the mechanism of nucleic acid transfer by liposome-DNA complexes, studies at the molecular level are required. To this end, scanning probe microscopy (SPM) is increasingly being used in the characterization of lipid layers, lipid aggregates, liposomes, and their complexes with nucleic acid molecules. The most attractive attributes of SPM are the potential to image samples with subnanometer spatial resolution under physiological conditions and provide information on their physical and mechanical properties. This review describes the application of scanning tunneling microscopy and atomic force microscopy, the two most commonly applied SPM techniques, in the characterisation of liposome-DNA complexes.

Design, synthesis and biological testing of a novel series of anti-inflammatory drugs.

Many of the non-steroidal anti-inflammatory drugs (NSAIDs) currently marketed produce severe gastro-toxic side effects. The benefits of producing NSAIDs without these side effects are obvious, particularly for patients requiring long-term therapy. The aim of this investigation was to produce novel NSAIDs, based on paracetamol, that exhibit little or no gastro-toxicity. The work covers design, synthesis and testing of 13 drug candidates. The analgesic and anti-inflammatory potencies of the drug candidates were measured using the mouse abdominal constriction assay and the carrageenan-induced rat paw oedema assay, respectively. The stomachs of the rats were examined post-mortem, to assess the gastro-toxicity of the drugs. Of the 13 compounds described herein, 11 were shown to possess analgesic activity at 2-10 times the potency of aspirin, while 8 demonstrated anti-inflammatory activity at 3-10 times the potency of aspirin. Significantly, all of the compounds showed very low gastro-toxicity when compared with aspirin. The results of this study indicate that it is possible to develop novel, potent NSAIDs based on the structure of paracetamol. These compounds have the advantage of demonstrating much lower gastro-toxicity than NSAIDs currently available. Drugs of this type may, in future, provide effective treatments for inflammatory disorders.

Deep lamellar keratoplasty with lyophilised tissue in the management of keratoconus.

AIMS: Data are presented on the use of deep lamellar keratoplasty (DLK) using lyophilised donor corneal tissue, in the management of patients with keratoconus (KC). METHOD: The results of DLK on 44 eyes (42 patients) are reported. The mean patient age was 29.8 years (range 10-56). Mean follow up was 25 months (range 6-100). In seven patients with mental handicap or severe mental illness, the collection of acuity and refractive data was limited. RESULTS: Perforation of Descemet's membrane (DM) occurred in nine cases (20%). A double anterior chamber formed in five cases, which resolved spontaneously in three patients. Persistent epithelial defects occurred in two cases, one of which necessitated replacement of the graft. The median postoperative uncorrected visual acuity was 6/36. The median corrected postoperative acuity was 6/9. Those with more than 1 year of follow up (n=25) had a significantly better acuity (p=0.015). This group achieved 6/12 or better in 80% (n=20) and 6/6 or better in 40% (n=10). The mean postoperative spherical error was +0.28 (SD 3.49) dioptres (D). The mean refractive cylinder was 3.85 (1.87) D. CONCLUSION: This detailed retrospective study of DLK for the treatment of patients with KC, with an average follow up of 2 years, highlights the advantages and disadvantages of this technique.

Synthesis and hydrolytic behaviour of glycerol-1,2-diibuprofenate-3-nitrate, a putative pro-drug of ibuprofen and glycerol-1-nitrate.

Nitroxylated derivatives of non-steroidal anti-inflammatory drugs appear to offer protection against the gastrotoxicity normally associated with non-steroidal anti-inflammatory drugs, ostensibly via local production of nitric oxide. A diester of ibuprofen and glycerol-1-mononitrate has been prepared via the condensation of ibuprofen with 3-bromopropan-1,2-diol, followed by silver-(I)-nitrate-mediated nitroxylation. The release of ibuprofen from this diester has been studied in a simulated gastric fluid model with direct analysis by reverse-phase HPLC, using an acetonitrile-water (80%:20%) mobile phase containing trifluoroacetic acid (0.005%). n-Propyl ibuprofen was found to undergo pH-dependent hydrolysis, ranging from negligible hydrolysis at pH 5 to 52% hydrolysis at pH 3, over a 2-h period in this model. The ibuprofen-glycerol mononitrate diester was subjected to the most vigorous model hydrolytic conditions and was found to undergo 50 % hydrolysis during the study period. This study shows that pro-drugs of ibuprofen and glycerol mononitrate can be obtained, and can undergo degradation to the parent drugs under conditions simulating those likely to be encountered in the stomach.

Corneal endothelial specular microscopy following deep lamellar keratoplasty with lyophilised tissue.

PURPOSE: To assess the corneal endothelial cell density following deep lamellar keratoplasty (DLK) carried out by air dissection and with the transplantation of lyophilised tissue. METHOD: Contact endothelial specular microscopy was carried out on a series of patients who had undergone DLK with a minimum of 1 year post-operative follow-up. RESULTS: Twenty eyes of 18 patients were examined at follow-up times ranging from 1 to 8 years post-operatively (mean 3 years). Indications for surgery were: keratoconus (n = 12), herpes simplex virus (HSV) keratitis (n = 1), herpes zoster ophthalmicus (HZO) keratitis (n = 1), lipid keratopathy (n = 2), lattice dystrophy (n = 1) and corneal scarring (n = 3). Overall, mean post-operative cell density was 2417 cells/mm2 (range 928-3656 cells/mm2). In eyes with pathological conditions not likely to have affected the endothelial cell density, such as keratoconus or lattice degeneration, the mean cell density was 2837 cells/mm2 (range 1030-3656 cells/mm2). CONCLUSIONS: In patients undergoing DLK for conditions such as keratoconus the post-operative cell density was at a normal level in the majority of cases. Cell loss through this surgical intervention thus appears generally to be small. The prospect for long-term survival of these grafts is good.

A prospective controlled study of a 10/0 absorbable polyglactin suture for corneal incision phacoemulsification.

PURPOSE: To evaluate the performance of a 10/0 monofilament absorbable polyglactin suture for temporal 5.2 mm corneal incision phacoemulsification. METHODS: A prospective randomised controlled study of 49 patients undergoing phacoemulsification with a sutured temporal 5.2 mm corneal section was conducted to compare the refractive results and complications of a 10/0 monofilament absorbable polyglactin suture with 10/0 nylon. RESULTS: Thirty-eight patients completed the study. There was no significant difference in induced astigmatism between the two groups. All absorbable sutures were intact at 1 week. Six weeks post-operatively the absorbable suture was still intact in 1 (6%) patient, present but broken in 4 (24%) and completely absent in 12 (70%) patients. All polyglactin sutures had been completely absorbed by 12 weeks. Absorption of the polyglactin sutures was associated with mild localised corneal haze in 3(18%) cases (p = 0.055). One of the 18 patients (6%) in the polyglactin suture group presented with iris prolapse associated with knot failure 1 week post-operatively. There was no significant difference in foreign body symptoms between the two groups. CONCLUSIONS: 10/0 polyglactin sutures maintain adequate tensile strength during the immediate post-operative period for small incision surgery and are associated with minimal induction of astigmatism. Their subsequent absorption obviates the need for routine suture removal. Suture absorption is well tolerated although in some cases a mild degree of local tissue reaction raises concern about possible mechanisms of absorption. The risk of knot failure may be reduced by an alternative suture tying technique. 10/0 monofilament polyglactin is an attractive option when a suture is required during small incision cataract surgery.

Intraocular lens implants and risk of endophthalmitis.

AIM: To investigate the possible association between the use of three piece foldable silicone polypropylene (SPP) intraocular lenses (IOLs) and an increased risk of postoperative endophthalmitis. METHODS: A retrospective analysis was conducted of all cases of postoperative endophthalmitis following phacoemulsification surgery in a single unit over a 3 year period. The incidence of postoperative endophthalmitis in eyes with SPP IOLs was compared with the incidence in eyes with single piece polymethylmethacrylate (PMMA) IOLs. RESULTS: 772 cataract extractions by phacoemulsification were performed. One (0.16%) of the 622 patients with PMMA IOLs developed endophthalmitis. Excluding one patient who had aplastic anaemia, five (3.33%) of 150 patients with SPP IOLs developed endophthalmitis. The relative risk for postoperative endophthalmitis associated with the use of the SPP IOL compared with the PMMA IOL was 20.1 (p = 0.015). CONCLUSION: This study adds further evidence to the concept that SPP IOLs can be a significant risk factor in the development of postoperative endophthamitis.

HLA antigens in lyophilised corneal lenticules.

PURPOSE: To investigate the presence of Human Leukocyte Antigens (HLA) in organ cultured corneas, in lyophilised epikeratophakia lenticules prepared by dry state lathing, and in lenticules subjected to prolonged rehydration in Balanced Salt Solution (BSS). METHODS: Twelve lenticules and 3 organ cultured corneas were studied for the presence of HLA-ABC (class I), and HLA-DR, -DQ, and -DP (class II), antigens using the immunoperoxidase technique. RESULTS: HLA-ABC antigens were detected in all lenticules. HLA-DR antigen was also detected in low concentrations in 6 out of 12 lenticules. HLA-DQ and -DP antigens were generally absent. Incubation in BSS for 24 and 48 hours did not change the expression of antigens. CONCLUSIONS: HLA-class I, and to some degree HLA-class II antigens, are present in the stroma of both organ cultured corneas, and in lyophilised lenticules. Incubation in BSS for up to 48 hours did not reduce the expression of antigens.

Systemic conditions in the Emergency Eye Clinic.

PURPOSE: A systemic disease may be diagnosed for the first time in patients who present to the eye department with ophthalmological manifestations. This study documents the pattern of systemic disorders which were encountered in an emergency eye clinic for the first time. SUBJECTS AND METHOD: Over a period of six months details of all patients who had a previously unknown systemic condition diagnosed in the Emergency Eye Clinic were recorded. Patients were seen in this clinic upon referral from other practitioners. All patients, with the exception of those with minor ophthalmic problems, were screened for hypertension and glycosuria. The patients were subsequently referred to their general practitioner or the appropriate hospital specialist for further management. RESULTS: During this period 1025 new patients visited the emergency eye clinic. Previously undiagnosed systemic disease was present in 86 patients (8.4%). The commonest disorders were neurological 35 (41%), cardiovascular 17 (20%), glycosuria 13 (15%) and respiratory 5 (6%). Visual disturbance and pain were the most frequent ocular presentation. In 54 patients (62.8%) the ocular condition was a direct manifestation of the systemic condition and in 32 patients (37.2%) the systemic disease was a coincidental finding. CONCLUSIONS: Awareness of ophthalmological manifestations of systemic disorders helps in the early diagnosis of these conditions and reduces the resulting morbidity and possibly mortality. This study illustrates the hidden role of the emergency eye clinic in identifying patients with an underlying systemic condition.

Fibrin glue: an alternative method of wound closure in glaucoma surgery.

PURPOSE AND METHODS: A commercially available tissue glue has been used to effect conjunctival wound closure after trabeculectomy. In four cases it was used in conjunction with sutures and in two cases alone to achieve watertight closure of the conjunctiva. A small transient leak was noted postoperatively in one case, and no case of flat anterior chamber occurred. RESULTS AND CONCLUSION: Intraocular pressure was controlled in all cases 3 months postoperatively. Tissue glue can be an effective method of achieving conjunctival wound closure in glaucoma surgery.