Guanine sulphinate is a major stable product of photochemical oxidation of DNA 6-thioguanine by UVA irradiation.

The DNA of patients taking the immunosuppressant and anticancer drugs azathioprine or 6-mercaptopurine contains 6-thioguanine (6-TG). The skin of these patients is selectively sensitive to ultraviolet A radiation (UVA) and they suffer an extremely high incidence of sunlight-induced skin cancer with long-term treatment. DNA 6-TG interacts with UVA to generate reactive oxygen species, which oxidize 6-TG to guanine sulphonate (G(SO3)). We suggested that G(SO3) is formed via the reactive electrophilic intermediates, guanine sulphenate (G(SO)) and guanine sulphinate (G(SO2)). Here, G(SO2) is identified as a significant and stable UVA photoproduct of free 6-TG, its 2'-deoxyribonucleoside, and DNA 6-TG. Mild chemical oxidation converts 6-TG into G(SO2), which can be further oxidized to G(SO3)-a stable product that resists further reaction. In contrast, G(SO2) is converted back to 6-TG under mild conditions. This suggests that cellular antioxidant defences might counteract the UVA-mediated photooxidation of DNA 6-TG at this intermediate step and ameliorate its biological effects. In agreement with this possibility, the antioxidant ascorbate protected DNA 6-TG against UVA oxidation and prevented the formation of G(SO3).

NCX3 knockout mice exhibit increased hippocampal CA1 and CA2 neuronal damage compared to wild-type mice following global cerebral ischemia.

There is uncertainty as to whether the plasma membrane Na(+)/Ca(2+)exchanger (NCX) has a neuroprotective or neurodamaging role following cerebral ischemia. To address this issue we compared hippocampal neuronal injury in NCX3 knockout mice (Ncx3(-/-)) and wild-type mice (Ncx3(+/+)) following global cerebral ischemia. Using a bilateral common carotid artery occlusion (BCCAO) model of global ischemia we subjected NCX3 knockout and wild-type mice to 17 and 15 minutes of ischemia. Following the 17 minute period of ischemia, wild-type mice exhibited approximately 80% CA1 neuronal loss and approximately 40% CA2 neuronal loss. In contrast, NCX3 knockout mice displayed >95% CA1 neuronal loss and approximately 95% CA2 neuronal loss. Following the 15 minute period of ischemia, wild-type mice did not exhibit any significant hippocampal neuronal loss. In contrast, NCX3 knockout mice displayed approximately 45% CA1 neuronal loss and approximately 25% CA2 neuronal loss. The results clearly demonstrate that mice deficient in the NCX3 protein are more susceptible to global cerebral ischemia than wild-type mice. Our findings suggest NCX3 has a positive role in maintaining neuronal intracellular calcium homeostasis following ischemia, and that when exchanger function is compromised neurons are more susceptible to calcium deregulation and cell death.

The role of the Na(+)/Ca(2+) exchanger (NCX) in neurons following ischaemia.

The Na(+)/Ca(2+) exchanger (NCX) is a bi-directional membrane ion transporter. Under normal conditions, the exchanger transports one calcium ion out of the cell and three sodium ions into the cell. This is known as the calcium exit, or "forward" mode. Under certain conditions, however, the exchanger can reverse and transport calcium ions into the cell (calcium entry mode). Because dysregulation of sodium and calcium homeostasis is an integral feature of ischaemic brain injury, the role of the NCX in neurons following ischaemia has been investigated using a number of in vitro and in vivo models. Studies using in vitro ischaemia-related models (hypoxia, glutamate) have produced conflicting results, with some showing that NCX activity is neuroprotective while others indicate that it is neurodamaging. The majority of in vivo studies using the focal cerebral ischaemia model indicate that blocking NCX activity is neurodamaging while increasing NCX activity is neuroprotective. We have reviewed the major in vitro and in vivo neuronal ischaemia-related NCX studies in an attempt to clarify the reason for the conflicting findings. The use of different ischaemia models and doubts as to the specificity of pharmacological NCX inhibitors and stimulators has contributed to the confusion over the role of the NCX in ischaemic brain injury. The development of NCX transgenic animals may help our understanding of the role of this ion exchanger in neurons following ischaemia and aid the development of an effective stroke treatment.

Novel DNA lesions generated by the interaction between therapeutic thiopurines and UVA light.

The therapeutic effect of the thiopurines, 6-thioguanine (6-TG), 6-mercaptopurine, and its prodrug azathioprine, depends on the incorporation of 6-TG into cellular DNA. Unlike normal DNA bases, 6-TG absorbs UVA radiation, and UVA-mediated photochemical damage of DNA 6-TG has potentially harmful side effects. When free 6-TG is UVA irradiated in solution in the presence of molecular oxygen, reactive oxygen species are generated and 6-TG is oxidized to guanine-6-sulfonate (G(SO3)) and guanine-6-thioguanine in reactions involving singlet oxygen. This conversion is prevented by antioxidants, including the dietary vitamin ascorbate. DNA G(SO3) is also the major photoproduct of 6-TG in DNA and it can be selectively introduced into DNA or oligonucleotides in vitro by mild chemical oxidation. Thermal stability measurements indicate that G(SO3) does not form stable base pairs with any of the normal DNA bases in duplex oligonucleotides and is a powerful block for elongation by Klenow DNA polymerase in primer extension experiments. In cultured human cells, DNA damage produced by 6-TG and UVA treatment is associated with replication inhibition and provokes a p53-dependent DNA damage response.

Leptomeningeal carcinomatosis: an unusual cause of sudden onset bilateral sensorineural hearing loss.

We report a 66-year-old woman who developed sudden-onset bilateral sensorineural deafness due to leptomeningeal carcinomatosis involving the vestibulocochlear nerves. The clinical and diagnostic features of leptomeningeal carcinomatosis are discussed.

Spinal cord ganglioglioma presenting as acute paraparesis.

A 17-year-old male presented with acute onset paraparesis in the lower limbs. Urinary retention was present and the patient required catheterisation. Clinical examination confirmed severe bilateral lower limb weakness and a sensory level at T8. Magnetic resonance imaging (MRI) revealed a haemorrhagic intramedullary tumour extending from T8 to the conus. Microsurgical excision of the tumour was performed and the patient made a good functional recovery. The histology of the tumour demonstrated a ganglioglioma of the spinal cord. Acute paraparesis has not previously been reported with a spinal cord ganglioglioma. We discuss the clinical, diagnostic and pathological features of spinal cord gangliogliomas.

Functioning paraganglioma of the thoracic spine: case report.

OBJECTIVE AND IMPORTANCE: Paragangliomas of the thoracic spine are rare. Previously described cases involved nonfunctioning tumors. This report documents the diagnosis and surgical treatment for a patient who presented with a functioning thoracic paraganglioma. CLINICAL PRESENTATION: A 53-year-old woman presented with a 10-month history of headaches, facial flushing, and palpitations associated with hypertension. Urinary catecholamine levels were markedly elevated. Magnetic resonance imaging and m-[(123)I]iodobenzylguanidine scans demonstrated an extradural tumor located within the T12 vertebra, with a significant paraspinal component. The neurological examination revealed mild hypesthesia in the right T12 dermatome. INTERVENTION: The patient underwent resection of the tumor after alpha-adrenergic receptor blockade. Grossly complete excision was achieved without neurological complications. Postoperatively, the patient was normotensive and exhibited catecholamine levels within the normal range. CONCLUSION: Functioning paragangliomas of the thoracic spine are rare lesions that are difficult to treat. Successful treatment requires careful surgical planning and expert pharmacological manipulation.

Enantioselection in peptide bond formation.

Selectivity in abiotic condensations of amino acids remains controversial and stereochemically little explored. We find that competitive activated couplings of N-acyl derivatives of glycine, alanine, valine, proline and phenylalanine with binary, ternary and quaternary mixtures of amides and esters of the same group of amino acids show little selectivity among the reactants, except with respect to configuration, where a consistent and significant preference for heterochiral outcomes, mostly > 80%, is observed. One possible explanation of this selectivity predicts a predisposition to homochiral coupling under conditions that would require the two carboxyl functions to be co-facial in the activated complex.