A pilot study to measure marks in children with cerebral palsy using a novel measurement template.

AIM: The primary aim of this pilot study was to trial a method of assessing bruises in a population of disabled children. If the method was found to be sufficiently robust it would be our intention to undertaking a more extensive observational study. BACKGROUND: Less is known about normal bruising patterns in children with disability than in those without. It is important that the method used to assess bruising is objective and repeatable. In an effort to define and improve repeatability, we employed a novel bruise measurement template which was printed onto transparent acetate sheets. METHOD: Twenty primary school age children, the majority of whom were non-ambulant and severely disabled with cerebral palsy, underwent full skin examination. The template was used to assess any bruises seen. A comparison was then made between measurements made by experienced paediatricians using the template and using a standard tape measure on a series of bruise images in 25 photographs. RESULTS: The majority of children in our pilot were found to have bruises, with one child having 6 and one 7 bruises. This comparative study showed that the two techniques had a very similar precision and that the template was easy to use. Greater precision would require a tighter measurement protocol, whether with a template or a tape measure. CONCLUSIONS: Further evaluation of the application of such a template would be worthwhile. We would suggest that our finding of some bruising in this population of disabled children is borne in mind whenever bruising is found in a non-ambulant child.

Potent 2'-amino-, and 2'-fluoro-2'-deoxyribonucleotide RNA inhibitors of keratinocyte growth factor.

Reiterative in vitro selection-amplification from random oligonucleotide libraries allows the identification of molecules with specific functions such as binding to specific proteins. The therapeutic usefulness of such molecules depends on their high affinity and nuclease resistance. Libraries of RNA molecules containing 2'amino-(2'NH2)- or 2'fluoro-(2'F)-2'-deoxypyrimidines could yield ligands with similar nuclease resistance but not necessarily with similar affinities. This is because the intramolecular helices containing 2'NH2 have lower melting temperatures (Tm) compared with helices containing 2'F, giving them thermodynamically less stable structures and possibly weaker affinities. We tested these ideas by isolating high-affinity ligands to human keratinocyte growth factor from libraries containing modified RNA molecules with either 2'NH2 or 2'F pyrimidines. We demonstrated that 2'F RNA ligands have affinities (Kd approximately 0.3-3 pM) and bioactivities (Ki approximately 34 pM) superior to 2'NH2 ligands (Kd approximately 400 pM and Ki approximately 10 nM). In addition, 2'F ligands have extreme thermo-stabilities (Tm approximately 78 degrees C in low salt, and specificities).

Nuclease-resistant nucleic acid ligands to vascular permeability factor/vascular endothelial growth factor.

BACKGROUND: Vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) is a potent inducer of new blood vessel growth (angiogenesis) that contributes to the pathology of many angiogenesis-associated disease states such as psoriasis, rheumatoid arthritis and cancer. Few molecular entities capable of binding to VPF/VEGF with high affinity and specificity have been described to date. RESULTS: Nuclease-resistant 2'-amino-2'-deoxypyrimidine nucleotide RNA (2'-aminopyrimidine RNA) ligands that bind to VPF/VEGF with high affinity have been identified by iterative rounds of affinity-selection/amplification from two independent random libraries. The sequence information that confers high affinity binding to VPF/VEGF is contained in a contiguous stretch of 24 nucleotides, 5'-CCCUGAUGGUAGACGCCGGGGUG-3' (2'-aminopyrimidine nucleotides are designated with italic letters). Of the 14 ribopurines in this minimal ligand, 10 can be substituted with the corresponding 2'-O-methylpurine nucleotides without a reduction in binding affinity to VPF/VEGF. In fact, the 2'-O-methyl substitution at permissive positions leads to a approximately 17-fold improvement in the binding affinity to VPF/VEGF. The higher affinity results from the reduction in the dissociation rate constant of the 2'-O-methyl-substituted RNA ligand from the protein compared to the unsubstituted ligand. The 2'-O-methyl-substituted minimal ligand, which folds into a bulged hairpin motif, is also more thermally stable than the unsubstituted ligand. Nuclease resistance of the ligand is further improved by the 2'-O-methyl substitutions and the addition of short phosphorothioate caps to the 3'- and 5'-ends. CONCLUSIONS: We have used the SELEX (systematic evolution of ligands by exponential enrichment) process in conjunction with post-SELEX modifications to define a highly nuclease-resistant oligonucleotide that binds to VPF/VEGF with high affinity and specificity.

Mammalian stanniocalcins and cancer.

Stanniocalcin (STC) is a glycoprotein hormone that is secreted by the corpuscle of Stannius, an endocrine gland of bony fish, and is involved in calcium and phosphate homeostasis. The related mammalian proteins, STC1 and STC2, are expressed in a wide variety of tissues. The ovaries have the highest level of STC1, and this increases during pregnancy and lactation. STC1 is present in breast ductal epithelium, and its expression is induced by BRCA1, a tumor suppressor gene that has an important role in breast and ovarian cancer. The expression of STC2 is induced by estrogen, and there is a positive correlation between the level of expression of estrogen receptor and expression of both STC1 and STC2 in breast cancer. This article reviews the data currently available regarding the mammalian STCs, and discusses the roles they may play in normal physiology and in breast and other cancers.

Subarachnoid hemorrhage due to late recurrence of a previously unruptured aneurysm after complete endovascular occlusion.

We present a case of subarachnoid hemorrhage attributed to rupture of an aneurysm 18 months after endovascular occlusion. The aneurysm was diagnosed after the patient had a seizure; however, there was no evidence of subarachnoid hemorrhage. Angiography at 6 months revealed a totally occluded aneurysm. This case illustrates that the long-term results of endovascular occlusion remain uncertain.

Multimodality MR imaging depiction of hemodynamic changes and cerebral ischemia in subarachnoid hemorrhage.

BACKGROUND AND PURPOSE: Subarachnoid hemorrhage (SAH) is a common and serious neurologic emergent condition. We tested the hypothesis that multimodality MR imaging depicts changes in cerebral blood flow SAH, before any surgical or endovascular intervention, and that the frequency of these changes increases with time after ictus. METHODS: We prospectively examined 37 patients with suspected SAH and three with symptoms of acute stroke but who subsequently had SAH. Routine CT and multimodality MR imaging were performed within 18 h of presentation. Standard MR imaging, diffusion-weighted MR imaging, time-of-flight MR angiography, and dynamic first-pass gadolinium-enhanced MR perfusion imaging were performed. Images were reviewed for abnormalities in cerebral blood flow, ischemia, and infarction. Nine patients did not have SAH at CT and CSF investigations. Of 31 patients with proved SAH, 13 were examined during the acute stage (within 4 d of ictus) and 18, during the subacute stage (4-14 d after ictus). RESULTS: MR imaging showed alteration in cerebral blood flow parameters in 16 of 31 patients before surgery or endovascular treatment. The frequency of blood flow changes and associated complications increased with worsening clinical grade and increasing time after ictus. CONCLUSION: Multimodality MR imaging provides information not available from CT in patients with SAH. MR imaging shows oligemic and ischemic areas in SAH before surgery or endovascular treatment. MR imaging is a simple noninvasive method of assessing cerebral blood flow and its complications in SAH. It can be performed in a clinical environment.

Noninvasive intraoperative monitoring of motor evoked potentials under propofol anesthesia: effects of spinal surgery on the amplitude and latency of motor evoked potentials.

We present the results of intraoperative monitoring of motor evoked potentials from 34 patients undergoing spinal surgery under total anesthesia with intravenously administered propofol. Intraoperative recording was performed with transcranial electrical stimulation. Two groups of patients were studied: 1) a control population of 26 patients undergoing lumbar discectomy for prolapsed intervertebral disc, all of whom had normal preoperative motor conduction; and 2) a population of 8 patients undergoing neurosurgical procedures for spinal tumor (5 patients) and spinal arteriovenous malformation (3 patients), all of whom had abnormal preoperative neurological signs and abnormal preoperative motor conduction. In the first group, electromyographic responses were recorded intraoperatively either from the 2nd dorsal interosseous muscle of the hand (5 patients) or from the 1st dorsal interosseous muscle of the foot (21 patients). In the second group, responses were recorded intraoperatively either from the 1st dorsal interosseous muscle of the foot (7 patients) or from the anterior tibial muscle (1 patient). Intraoperative monitoring of motor function was successful in 88.5% of the patients in the control group. Propofol anesthesia caused a reduction in response amplitude to 7% of baseline values obtained from conscious relaxed subjects. Intraoperative monitoring was successful in 87% of the patients in the pathological group. We observed significant changes in both amplitude (greater than 50%) and/or onset latency (greater than 3 ms) from the intraoperative baseline that indicated either improvement (3 patients) or deterioration (2 patients) in motor conduction within minutes of surgical maneuvers anticipated to alter spinal cord function. Only permanent complete loss of intraoperative motor conduction (1 patient) correlated with a significant change in the postoperative neurological state.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of nitrous oxide on motor evoked potentials recorded from skeletal muscle in patients under total anesthesia with intravenously administered propofol.

The effect of nitrous oxide (N2O) on motor evoked potentials (MEPs) recorded from human subjects under total intravenous anesthesia with propofol (2,6-diisopropylphenol) was studied. MEPs were recorded from the 1st dorsal interosseous muscle of the foot in nine subjects; in two of these, simultaneous recordings were made from the 2nd dorsal interosseous muscle of the hand and from the deltoid muscle. Single transcranial electrical stimuli were used in recording the MEPs. The effects of N2O were studied at concentrations from 20 to 70%. Increasing concentrations of N2O caused a progressive increase in onset latency and a fall in the peak-to-peak amplitude of the MEPs recorded from the foot. Latency values showed a significant increase above the baseline at concentrations of N2O greater than 20% (P values, 0.05-0.005). The response amplitude showed a significant decrease from the baseline at concentrations of N2O greater than 50% (P values, 0.05-0.005). The 2nd dorsal interosseous muscle of the hand demonstrated a pattern of sensitivity to N2O similar to that of the 1st dorsal interosseous muscle of the foot. The onset latency and initial peak-to-peak amplitude of the deltoid muscle were insensitive to N2O at the concentrations used. We conclude that N2O can be used as an anesthetic adjunct without a significant deleterious effect on MEPs during intraoperative monitoring in patients under propofol anesthesia, providing concentrations are maintained below 50%.

Motor functional MRI for pre-operative and intraoperative neurosurgical guidance.

Functional MRI (fMRI) may provide a means of locating areas of eloquent cortex that can be used to guide neurosurgeons in their quest to maximize intracerebral tumour resection whilst minimizing post-procedural neurological deficits. This work aimed to develop and provide an initial assessment of such a technique. 19 patients with mass lesions close to the primary motor cortex underwent fMRI at 1.5T. A single shot echo planar technique was used to acquire data corresponding to right and left hand movement. Resultant activation maps were used to aid pre-surgical planning. Data was used in conjunction with an intraoperative navigation system in 13 cases. Activation was attributed to primary motor, primary somatosensory or supplementary motor cortex in 17 of 19 subjects. No permanent changes in motor deficit were detected post surgery. The additional information provided by fMRI, particularly when incorporated into a neuronavigation guided craniotomy, was deemed highly valuable to the neurosurgeon as it enabled safe resection of tumour in anatomical locations previously deemed to be too high risk for safe resection using conventional (non-fMRI-guided) technique. This observation is reinforced by the fact that no patients suffered permanent neurological deficit after radical tumour debulking (surgical estimates >90% tumour resection).

A case of pneumocephalus secondary to VP shunt in a patient with acoustic neuroma.

Pneumocephalus is an uncommon complication of ventriculo-peritoneal (VP) shunts. We present a case of pneumocephalus secondary to a VP shunt in a patient with an acoustic neuroma. The site of the cerebrospinal fluid fistula was found to be into an extensive petrous apex air cell system due to tumour erosion of the internal auditory canal.

Diffuse leptomeningeal spread of pleomorphic xanthoastrocytoma.

Diffuse meningeal spread and dural infiltration by pleomorphic xanthoastrocytoma occurring after initial diagnosis of a left frontotemporal lesion is reported. The unusual pattern of spread and aggressive course for a pleomorphic xanthoastrocytoma are discussed.

Lymphoblastic leukaemia presenting as a carotid-cavernous fistula.

A carotid-cavernous fistula is a life-threatening condition characterised by an abnormal communication between the carotid arterial vessels and the cavernous venous system. Although these fistulae can arise spontaneously, they mainly occur after trauma, especially road traffic accidents, falls and penetrating cranial or orbital injuries. The mainstay of treatment involves endovascular embolization, but in those patients where this is not possible or where embolization fails, direct surgical intervention and ligation of the artery may be necessary. Here we describe an interesting case of a suspected carotid-cavernous fistula which turned out to be cavernous sinus syndrome secondary to lymphoma.

Subtypes of oligodendroglioma defined by 1p,19q deletions, differ in the proportion of apoptotic cells but not in replication-licensed non-proliferating cells.

Oligodendrogliomas may be divided into those with deletion of chromosomes 1p and 19q (Del+), and those without (Del-). Del+ tumours show better survival and chemoresponsiveness but the reason for this difference is unknown. We have investigated whether these subgroups differ in (a) apoptotic index, (b) the proportion of cells licensed for DNA replication but not in-cycle, and (c) the relative length of G1-phase. Fluorescence in situ hybridisation with probes to 1p and 19q was used to determine the deletion status of 54 oligodendrogliomas, including WHO grades II and III. The apoptotic index was determined using counts of apoptotic bodies. Replication-licensed non-proliferating cells were determined from the Mcm2 minus Ki67 labelling index, whilst the geminin to Ki67 ratio was used as a measure of the relative length of G1. Del+ oligodendrogliomas showed a higher apoptotic index than Del- tumours (P=0.037); this was not accounted for by differences in tumour grade or in proliferation. There were no differences in the Mcm2-Ki67 index or in the geminin/Ki67 ratio between the subgroups, but grade III tumours showed a higher proportion of licensed non-proliferating cells than grade II tumours (P=0.001). An increased susceptibility to apoptosis in oligodendrogliomas with 1p+/-19q deletion may be important in their improved clinical outcome compared to Del- tumours.

Extra-cranial metastasis of glioblastoma multiforme presenting as acute parotitis.

We present an unusual case of extracranial metastasis of glioblastoma multiforme (GBM) to the parotid gland and cervical lymph nodes. The patient had previously undergone two craniotomies to debulk a left frontal GBM, followed by radiotherapy. After the second craniotomy, while waiting for chemotherapy, the patient was re-admitted with a short history of a painful swelling of his left parotid gland. The initial diagnosis was infective parotitis; however, as there was no improvement with broad-spectrum antibiotics, CT was undertaken, which revealed a mass in the parotid gland with a necrotic centre and enlarged cervical lymph nodes. Parotid gland biopsy revealed a parotid GBM metastasis. This case illustrates how GBM behaves in an aggressive manner even outside the CNS. A brief review of the literature and of the theories, which might explain the extra-neural metastasis of this tumour is also presented.

How are children's growth problems diagnosed?

Members of the Child Growth Foundation were surveyed to ascertain how children's growth problems were diagnosed. For many (46%) it was the parents who first expressed a concern about the child's growth. Routine height monitoring detected only 8%. Parents consulted a doctor promptly when worried, but there were often delays between the first medical consultation and referral to a hospital consultant. Many parents reported that their concerns were not taken seriously.

The impact of continuous electrophysiological monitoring on preservation of the facial nerve during acoustic tumour surgery.

Continuous electromyographic (EMG) activity and responses to electrical stimulation of the facial nerve were monitored intraoperatively in 35 patients undergoing primary removal of acoustic neuroma. The rate of anatomical and immediate functional preservation of the facial nerve of these patients was compared with a matched population where a facial nerve stimulator and monitoring of facial muscle contraction by the anaesthetist only was employed. Overall, 94% (n = 33) of facial nerves were preserved anatomically in patients with continuous EMG monitoring, compared with 64% (n = 89) of patients (n = 139) without continuous monitoring. Amongst the 33 cases with anatomical facial nerve preservation, 64% (n = 21) had immediate functional preservation. Immediate functional preservation was only achieved in 39% (n = 35) of anatomically preserved facial nerves (n = 89) where conventional monitoring was used.

Aneurysmal bone cyst of the orbit: a case report and review of literature.

Aneurysmal bone cyst is a benign fibroosseous lesion which rarely occurs in the orbit. We report on a 7-year-old girl with aneurysmal bone cyst of the orbit who presented with painless proptosis and diplopia. Optic nerve compression resulted in field loss and delayed visual evoked potentials. Radiological and histological features are discussed. The lesion was excised via a frontal craniotomy and the orbital roof reconstructed with a prefabricated titanium plate. Post-operatively a rapid resolution of the proptosis and diplopia followed. Previous reported cases of this rare entity in the orbit are also reviewed.

Needs of disabled children and their families.

In the new NHS those who provide services for disabled children need to measure and demonstrate their effectiveness, but there are no easily available outcome measures for use by child development centres and teams. The development of an alternative approach, using a series of statements of good practice, is described. Parents of children with cerebral palsy were asked to participate in semistructured interviews, to ascertain the value and relevance of these quality statements. Parents were most concerned about the standard of news breaking and early follow up, the sharing of information, and the supply and repair of equipment. The findings were used to modify the quality checklist and it is proposed that this should form the basis of a "charter for disabled children and their families'.

Inhibition of receptor binding by high-affinity RNA ligands to vascular endothelial growth factor.

The proliferation of new blood vessels (angiogenesis) is a process that accompanies many pathological conditions including rheumatoid arthritis and solid tumor growth. Among angiogenic cytokines that have been identified to date, vascular endothelial growth factor (VEGF) is one of the most potent. We used SELEX [systematic evolution of ligands by exponential enrichment; Tuerk, C., & Gold, L. (1990) Science 249, 505-510] to identify RNA ligands that bind to VEGF in a specific manner with affinities in the low nanomolar range. Ligands were selected from a starting pool of about 10(14) RNA molecules containing 30 randomized positions. Isolates from the affinity-enriched pool were grouped into six distinct families on the basis of primary and secondary structure similarities. Minimal sequence information required for high-affinity binding to VEGF is contained in 29-36-nucleotide motifs. Binding of truncated (minimal) high-affinity ligands to VEGF is competitive with that of other truncated ligands and heparin. Furthermore, truncated ligands from the six ligand families inhibit binding of [125I]VEGF to its cell-surface receptors. Oligonucleotide ligands described here represent an initial set of lead compounds in our ongoing effort toward the development of potent and specific VEGF antagonists.

Separation of growth-promoting activity for human muscle cells from fetuin.

Whether the growth-promoting activity of Pedersen fetuin is due to fetuin itself or to a contaminant(s) has been a long-standing puzzle. The possibility that the growth-promoting activity of Pedersen fetuin for human muscle satellite cells (HMSC) could be caused by some other component of fetal bovine serum (FBS) that remained in the fetuin as a contaminant has been investigated. One liter of FBS was first precipitated with 50% saturated ammonium sulfate, which leaves the serum albumin in solution, and then with 25% polyethylene glycol, which leaves the fetuin in solution, to generate a fraction 50 PEG 2x that was enriched 11-fold in growth-promoting activity for HMSC, with 68% recovery of total activity. Further purification with FPLC anion exchange chromatography achieved 99-fold enrichment of the activity with 30% overall recovery. The activity is heat labile and pH sensitive, suggesting that it is of protein nature, and the size of the activity is above 70 kDa. SDS-PAGE of the most active fractions shows that they are virtually free of fetuin. Thus, although the active fractions are not homogeneous, these studies demonstrate that the growth-promoting activity for HMSC can be fully separated from fetuin.