Synthesis of a Naphthalocyanine-Like Dye: The First Report on Zn(II)-1,6-methano[10]annulenecyanine.

The synthesis of the new dye 1,6-methano[10]annulenecyanine is described. For this purpose, the 3,4-dicyano-1,6-methano[10]annulene and 3,4-carboxyimide-1,6-methano[10]annulene buildings blocks were synthesized in six to eight steps. In both cases, these building blocks were then cyclotetramerized to furnish a new Zn(II)-1,6-methano[10]annulenecyanine which presents a strong red-shifted absorption band at 800 nm and high solubility in common organic solvents.

Understanding the photophysical properties of rhenium(I) compounds coordinated to 4,7-diamine-1,10-phenanthroline: synthetic, luminescence and biological studies.

In the present study, the photophysical properties and preliminary time-dependent density functional theory (TD-DFT) data of new rhenium(i) polypyridyl compounds, fac-[Re(L)(Am2phen)(CO)3]0/+, where Am2phen = 4,7-diamine-1,10-phenanthroline and L = Cl and ethyl isonicotinate (et-isonic), provided new insights into excited-state deactivation through an unusual inversion between two metal-to-ligand charge-transfer excited states. In addition, their cellular uptake using breast cancer (MCF-7) and melanoma (SkMel-147 and SkMel-29) cell lines and bioactivity were investigated and their cell-killing mechanism and protein expression were also studied. Preliminary TD-DFT results showed that both compounds exhibited a strong and broad absorption band around 300-400 nm which corresponds to a combination of ILAm2phen and MLCTRe→Am2phen transitions, and a strong contribution of charge transfer transition MLCTRe→et-isonic for fac-[Re(et-isonic)(Am2phen)(CO)3]+ is also observed. In contrast to typical Re(i) polypyridyl complexes, the substitution of Cl with the et-isonic ligand showed a bathochromic shift of the emission maxima, relatively low emission quantum yield and fast lifetime. Photophysical investigation of the fac-[ReCl(et-isonic)2(CO)3] compound provided meaningful information on the excited state manifold of the fac-[Re(L)(Am2phen)(CO)3]0/+ complexes. As shown in the absorption profile, a remarkable inversion of the lowest-lying excited state takes place from the usually observed MLCTRe→Am2phen to the unusual MLCTRe→et-isonic. The lipophilicity of the positive-complex was higher than that of the non-charge compound and the same trend for the activity against cells was observed, in the absence of light. In addition, flow cytometry and Western Blot analyses showed an overexpression of pro-caspase-9, suggesting a caspase proteolytic cascade through an intrinsic-pathway apoptosis mechanism. The photophysical properties of these compounds reported herein provide new fundamental insights into the understanding of substituent groups on polypyridyl ligands which are relevant to practical development.

Pharmacological Approaches to the Management of Secondary Progressive Multiple Sclerosis.

It is well recognised that the majority of the impact of multiple sclerosis (MS), both personal and societal, arises in the progressive phase where disability accumulates inexorably. As such, progressive MS (PMS) has been the target of pharmacological therapies for many years. However, there are no current licensed treatments for PMS. This stands in marked contrast to relapsing remitting MS (RRMS) where trials have resulted in numerous licensed therapies. PMS has proven to be a more difficult challenge compared to RRMS and this review focuses on secondary progressive MS (SPMS), where relapses occur before the onset of gradual, irreversible disability, and not primary progressive MS where disability accumulation occurs without prior relapses. Although there are similarities between the two forms, in both cases pinpointing when PMS starts is difficult in a condition in which disability can vary from day to day. There is also an overlap between the pathology of relapsing and progressive MS and this has contributed to the lack of well-defined outcomes, both surrogates and clinically relevant outcomes in PMS. In this review, we used the search term 'randomised controlled clinical drug trials in secondary progressive MS' in publications since 1988 together with recently completed trials where results were available. We found 34 trials involving 21 different molecules, of which 38% were successful in reaching their primary outcome. In general, the trials were well designed (e.g. double blind) with sample sizes ranging from 35 to 1949 subjects. The majority were parallel group, but there were also multi-arm and multidose trials as well as the more recent use of adaptive designs. The disability outcome most commonly used was the Expanded Disability Status Scale (EDSS) in all phases, but also magnetic resonance imaging (MRI)-measured brain atrophy has been utilised as a surrogate endpoint in phase II studies. The majority of the treatments tested in SPMS over the years were initially successful in RRMS. This has a number of implications in terms of targeting SPMS, but principally implies that the optimal strategy to target SPMS is to utilise the prodrome of relapses to initiate a therapy that will aim to both prevent progression and slow its accumulation. This approach is in agreement with the early targeting of MS but requires treatments that are both effective and safe if it is to be used before disability is a major problem. Recent successes will hopefully result in the first licensed therapy for PMS and enable us to test this approach.

Technical assessment of whole body angiography and cardiac function within a single MRI examination.

AIM: To evaluate a combined protocol for simultaneous cardiac MRI (CMR) and contrast-enhanced (CE) whole-body MR angiography (WB-MRA) techniques within a single examination. MATERIALS AND METHODS: Asymptomatic volunteers (n = 48) with low-moderate risk of cardiovascular disease (CVD) were recruited. The protocol was divided into four sections: (1) CMR of left ventricle (LV) structure and function; (2) CE-MRA of the head, neck, and thorax followed by the distal lower limbs; (3) CMR LV "late gadolinium enhancement" assessment; and (4) CE-MRA of the abdomen and pelvis followed by the proximal lower limbs. Multiple observers undertook the image analysis. RESULTS: For CMR, the mean ejection fraction (EF) was 67.3 ± 4.8% and mean left ventricular mass (LVM) was 100.3 ± 22.8 g. The intra-observer repeatability for EF ranged from 2.1-4.7% and from 9-12 g for LVM. Interobserver repeatability was 8.1% for EF and 19.1 g for LVM. No LV delayed myocardial enhancement was observed. For WB-MRA, some degree of luminal narrowing or stenosis was seen at 3.6% of the vessel segments (involving n = 29 of 48 volunteers) and interobserver radiological opinion was consistent in 96.7% of 1488 vessel segments assessed. CONCLUSION: Combined assessment of WB-MRA and CMR can be undertaken within a single examination on a clinical MRI system. The associated analysis techniques are repeatable and may be suitable for larger-scale cardiovascular MRI studies.

Assessing dalfampridine efficacy in the physician's office.

Dalfampridine (extended release 4-aminopyridine) is shown in three recent randomised controlled trials to improve walking speed in people with multiple sclerosis; however, the trial literature makes it clear that dalfampridine is effective in only a subset of patients. For the neurologist working in an everyday physician's office, a key question arises: How to distinguish the few who experience a meaningful clinical benefit, from the many who do not? This question has not yet been adequately addressed in the available literature.

Diagnosing cardiovascular disease from the perspective of the brain and inflammation.

Numerous lifestyle, emotional, and biological factors have been identified as risk factors for heart disease. These include socioeconomic status, early childhood and intimate partner abuse, disruption of sleep patterns, lack of exercise, and unhealthy food choices. Genetic and epigenetic factors are also critical components of the equation. A common denominator that links directly or indirectly all of these factors is inflammation. In some instances, the production of inflammatory molecules may precipitate the illness, while in others they may be produced in response to the underlying cause. Regardless of whether through direct or indirect means, inflammation contributes to the gradual loss of cellular energy substrates, which culminates in impaired diastolic performance. For that reason, to refer to a failure of the cardiovascular system as heart or cardiovascular disease lessens the potentially important contribution of a myriad of other factors. This article begins with the premise that impaired cardiac functioning is more than a heart disorder. An argument will be made that impaired cardiac functioning can also be an economic, behavioral, and/or emotional disorder, which subsequently gives rise to a metabolic failure. Therefore, a multi-systems approach should be taken to identify prior to the onset of damage biological and non-biological predictors of impending heart disease.

New columnar Zn-phthalocyanine designed for electronic applications.

Columnar liquid crystals are composed of disk-shaped aromatic molecules surrounded by flexible side chains, where molecules self-assemble in columns and thereby form large surface-oriented domains. These systems are known for their good charge and exciton transport along the columns, with mobilities approaching those of aromatic single crystals. Such semiconducting materials are promising for devices applications, since the output efficiency can be tuned by properly aligning columns. In the work presented here, the synthesis and characterization of a new Zn-phthalocyanine (ZnPc) is described which exhibits remarkable properties, such as hexagonal columnar order, achieved by cooling down from the isotropic phase to room temperature. Such order was confirmed by optical microscopy and X-ray diffraction experiments. Diodes were constructed using spin-coated films, and the conductive properties were investigated by current versus voltage analysis, where mobilities of 10(-3) and 10(-2) cm(2)/(V s) were obtained for the nonannealed and annealed films, respectively.

Quantitative analysis of cardiac left ventricular variables obtained by MRI at 3 T: a pre- and post-contrast comparison.

Short-axis cine images are acquired during cardiac MRI in order to determine variables of cardiac left ventricular (LV) function such as ejection fraction (EF), end-diastolic volume (EDV), end-systolic volume (ESV), stroke volume (SV) and LV mass. In cardiac perfusion assessments this imaging can be performed in the temporal window between first pass perfusion and the acquisition of delayed enhancement images in order to minimise overall scanning time. The objective of this study was to compare pre- and post-contrast short-axis LV variables of 15 healthy volunteers using a two-dimensional cardiac-gated segmented cine true fast imaging with steady state precession sequence and a 3.0 T MRI unit in order to determine the possible effects of contrast agent on the calculated cardiac function variables. Image analysis was carried out using semi-automated software. The calculated mean LV mass was lower when derived from the post-contrast images, relative to those derived pre-contrast (102 vs 108.1 g, p<0.0001). Small but systematic significant differences were also found between the mean pre- and post-contrast values of EF (69.4% vs 68.7%, p<0.05), EDV (142.4 vs 143.7 ml, p<0.05) and ESV (44.2 vs 45.5 ml, p<0.005), but no significant differences in SV were identified. This study has highlighted that contrast agent delivery can influence the numerical outcome of cardiac variables calculated from MRI and this was particularly noticeable for LV mass. This may have important implications for the correct interpretation of patient data in clinical studies where post-contrast images are used to calculate LV variables, since LV normal ranges have been traditionally derived from pre-contrast data sets.

A clinical MRI investigation of the relationship between kidney volume measurements and renal function in patients with renovascular disease.

Recent improvements in MR image acquisition and post-processing techniques have allowed quantitative kidney volume measurements to be derived from patient studies. These morphological indices can provide "snapshot" assessments that may be related to kidney function. The study objective was to measure cortical and total kidney volumes in patients with renovascular disease (RVD) using contrast-enhanced MR angiography (CE-MRA) in order to assess the reproducibility of the technique and to investigate associations between volumes and renal function as measured by glomerular filtration rate (GFR) calculations. 50 patients with RVD were scanned using CE-MRA. Kidney lengths, volumes and renal artery stenoses (RAS) were evaluated, and GFR was calculated using clinical formulae and nuclear medicine isotope renography. Mean MRI kidney lengths were 10.3+/-0.2 cm, and mean MRI volumes were 74.9+/-3.6 cm3 (cortical) and 128.5+/-5.3 cm3 (total). Kidneys supplied by moderately stenosed arteries had enlarged lengths and volumes, whilst those supplied by severely stenosed arteries had significantly smaller lengths (p<0.001) and volumes (p<0.001). There was a clear association between MRI cortical volume and GFR (r = 0.74, p<0.001, n = 48), but less so between kidney length and GFR (r = 0.54, p<0.001, n = 48). For individual patients, left/right cortical volume differences were small provided that severe RAS was not present, but large left/right volume differences and a GFR reduction were noted when severe RAS was present. The cortical volume distribution provides a useful single-timepoint indication of kidney function as defined by GFR, with no additional data acquisition required other than that of standard CE-MRA examination.

Outcome predictors and complications in the management of intradural spinal tumours.

The results of the management of 115 patients with intradural spinal tumours are presented. Data was collected retrospectively from the case notes. Tumours were categorized as intramedullary or extramedullary for statistical analysis. Meningioma, schwannoma and ependymoma accounted for 70% of tumours. Complete macroscopic excision was achieved in 84% of extramedullary and 54% of intramedullary tumours. There were two post-operative deaths, one of which was secondary to methacillin-resistant staphylococcus aureus (MRSA) meningitis. Cerebrospinal fluid leak (10%) and meningitis (7%) were the commonest complications. Ninety-six percent of patients with extramedullary tumours improved or remained unchanged on the Frankel scale. In the intramedullary group, 82% remained unchanged or improved after treatment. Pre-operative functional status was a predictor of good post-operative function for intra- and extramedullary tumours and for intramedullary tumours a good post-operative Frankel score predicted long-term survival.

The role of the PTPRC (CD45) mutation in the development of multiple sclerosis in the North West region of the United Kingdom.

BACKGROUND: A point mutation in protein tyrosine phosphatase receptor, type c polypeptide (PTPRC) has been associated with familial multiple sclerosis. This CG mutation at position 77 of exon 4 results in altered expression of CD45 isoforms on immune cells. OBJECTIVE: To study the incidence of PTPRC mutations in subjects with multiple sclerosis in the North West region of the United Kingdom. METHODS: Affected and unaffected subjects from five pedigrees with familial multiple sclerosis, 330 non-familial cases of multiple sclerosis, and 197 controls were studied. Genomic DNA was amplified using CD45IE34 and CD45IE44 primers, digested with Mspl, and run on an agarose gel. Polymerase chain reaction products were sequenced to exclude any other mutations. RESULTS: No PTPRC exon 4 genomic mutations were seen in any of the five families. In the non-familial cases the incidence of mutation was 4.1% in 197 controls and 5.1% in 330 multiple sclerosis patients. No significant association was found in this study with this mutation and disease susceptibility, sex, or an extended disability scale score of < 5.5. CONCLUSIONS: This candidate does not appear to influence the development of familial multiple sclerosis in this population. The negative result could arise from a type II error owing to the number of families and non-familial cases screened. Alternatively it might suggest that the contribution of the PTPRC mutation depends upon the genetic background.

Microglia-derived IGF-2 prevents TNFalpha induced death of mature oligodendrocytes in vitro.

Insulin-like growth factor-2 (IGF-2) present in media conditioned by non-activated and interferon gamma (IFN gamma)-treated microglia reduces galactocerebroside(+) (GalC) oligodendrocyte apoptosis in cultures derived both from the CG4 cell line and primary rat cortices. Microglia-derived IGF-2 also acts in each culture system to block GalC(+) oligodendrocyte toxicity resulting from soluble microglial-derived tumour necrosis factor alpha (TNF alpha). IGF-2 inhibits TNF alpha-induced c-Jun kinase (JNK) activation of the CG4 cell line. Microglial activation results in the release of soluble factors that are potentially toxic to oligodendrocytes but this may be offset by the production of soluble factors that protect these vulnerable cells. Allowing for extrapolation of these in vitro findings to intact tissue, our observations suggest one mechanism for limiting bystander damage in the context of inflammatory brain disease.

Nonactivated microglia promote oligodendrocyte precursor survival and maturation through the transcription factor NF-kappa B.

We demonstrate a role for nonactivated rat microglia in the survival and maturation of oligodendrocyte precursor cells (OPCs). Media conditioned by nonactivated microglia increase the number of surviving galactocerebroside(+) (GalC(+)) oligodendrocytes in vitro at 48 h by inhibiting the apoptosis of OPCs and stimulating their maturation to GalC+ oligodendrocytes. These effects are not observed with medium conditioned by microglia activated with interferon-gamma (IFN-gamma). Conditioned medium from nonactivated microglia is associated with upregulation in OPCs of nuclear factor of kappa binding (NF-kappa B) p65 subunit. The use of antisense to the inhibitor of kappa binding (I kappa B) induces p65 subunit activation in OPCs and, in common with medium conditioned by nonactivated microglia, also inhibits OPC apoptosis and promotes cell maturation. Anti-platelet-derived growth factor (PDGF) antibody abolishes this effect even though PDGF-A chain is expressed at similar levels within both nonactivated and IFN-gamma-activated microglia and both conditioned media have similar levels of PDGF-A chain bioactivity. However, only conditioned medium from nonactivated microglia recruit phosphatidyl-3-inositol (PI-3) kinase to the PDGF-alpha receptor and synergise with endogenous PDGF-A chain to increase NF-kappa B activation. These results suggest that, dependent on their state of activation, microglia produce soluble factors that promote oligodendrocyte development through an effect on the PDGF-alpha receptor-signalling pathway.

Inhibition of tumour necrosis factor-alpha (TNFalpha)-induced NF-kappaB p52 converts the metabolic effects of microglial-derived TNFalpha on mouse cerebellar neurones to neurotoxicity.

Activated microglia are implicated in the injury of neurones and macroglia both in vitro and in vivo. Here, we demonstrate that media conditioned by interferon-gamma treated microglia initially impair the metabolism of mouse cerebellar neurones grown in serum-free conditions without inducing cell death. Metabolic effects include inhibition of the ability of mitochondria to reduce 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and cytochrome oxidase activity. These effects are blocked by antibodies to tumour necrosis factor-alpha (TNFalpha), a cytokine produced by microglial activation, and they are not reproduced by media conditioned by resting microglia. The metabolic effects are evident for up to 24 h in vitro. More prolonged exposure, up to 48 h, results in TNFalpha dependent neuronal death as previously observed. Between 2 and 48 h TNFalpha present in media conditioned by interferon-gamma treated but not resting microglia is associated with nuclear factor kappa B (NF-kappaB) consensus sequence binding in paired mouse cerebellar neuronal cultures without affecting activation of the signal transducer and activator of transcription (STAT) transcription factor. Neuronal death can be accelerated by peptide blockade of the nuclear transport of NF-kappaB p52 subunit during exposure of cerebellar neurones to medium from interferon-gamma treated microglia. This toxicity is blocked by anti-TNFalpha antibody. Soluble factors released by activated microglia therefore contribute to neuronal dysfunction that is initially reversible but may culminate in neurotoxicity. Characterizing and manipulating these events in vivo theoretically provides an opportunity for neuroprotection in selected diseases affecting the central nervous system.

Peripheral inflammation increases the capsaicin sensitivity of dorsal root ganglion neurons in a nerve growth factor-dependent manner.

Inflammation results in a local increase in nerve growth factor production which potentially can modify the properties of nerve growth factor-responsive sensory neurons innervating the inflamed tissue. The sensitivity of primary sensory neurons to the neurotoxin capsaicin is regulated in vitro by nerve growth factor and we have now investigated the effect of complete Freund's adjuvant-induced inflammation on the capsaicin sensitivity of adult rat sensory neurons. Dorsal root ganglion neurons innervating inflamed tissue were identified in vivo by retrograde labelling with the dye Fast Blue. Neuronal capsaicin sensitivity was measured in vitro with a quantitative cobalt-uptake densitometric technique, and was shown to increase significantly five days after inflammation. This increase in sensitivity was dependent on nerve growth factor as it could be inhibited by systemic treatment with nerve growth factor neutralizing antibodies. The enhanced capsaicin sensitivity that results from Freund's adjuvant injection may contribute to inflammatory hyperalgesia.

Comparison of C1q-receptors on rat microglia and peritoneal macrophages.

A comparison of the expression and ligand specificity of the C1q (first complement component) receptor on rat microglia and peritoneal macrophages was made. This revealed that radiolabelled C1q was competed from the peritoneal macrophages with intact C1q, and additively displaced by calf-skin collagen and purified C1q globular heads, suggesting the presence of at least two receptors. This was in contrast to microglia, where radiolabelled C1q was displaced with intact C1q and to a modest degree with collagen, but not with globular heads. Taken together, this implies that under these conditions, peritoneal macrophages and microglia both express a C1q receptor which binds to the collagen-like region, and that peritoneal macrophages additionally express a molecule which binds to the globular head of C1q. Analysis of the ligand bound by these cells reflected the differences observed in the competitive binding experiments, with the novel identification of naturally-occurring peptides from the globular head of C1q bound to the peritoneal macrophages, but not the microglia.

A neurological rehabilitation unit: audit of activity and outcome.

A clinical audit was carried out to determine the impact of multidisciplinary rehabilitation in a specialist neurorehabilitation unit, and to demonstrate how outcome measurement can be incorporated into routine clinical audit. The study describes and interprets the results of one year's activity and outcome in a neurorehabilitation unit. A total of 138 patients were admitted to the 18 bedded unit between April 1994 and March 1995. The main outcome measures were: length of inpatient stay, admission and discharge destination, disability as measured by the Barthel Index and Functional Independence Measure, handicap as measured by the Environmental Status Scale and the Handicap Assessment Scale, and the time spent undertaking the audit. Improvement in disability was demonstrated in 112 (83%) patients and in handicap in 89 (66%) patients. The time taken to analyse the data on a quarterly basis was reduced from 20 hours for the first quarter to 4.5 hours for the last quarter. The results show that multidisciplinary inpatient neurorehabilitation leads to functional improvement in the majority of neurologically impaired patients. Outcome measurement and data collection can be incorporated into routine clinical practice once a sound methodology has been established.