DNA methylation in ELOVL2 and C1orf132 correctly predicted chronological age of individuals from three disease groups.

Improving accuracy of the available predictive DNA methods is important for their wider use in routine forensic work. Information on age in the process of identification of an unknown individual may provide important hints that can speed up the process of investigation. DNA methylation markers have been demonstrated to provide accurate age estimation in forensics, but there is growing evidence that DNA methylation can be modified by various factors including diseases. We analyzed DNA methylation profile in five markers from five different genes (ELOVL2, C1orf132, KLF14, FHL2, and TRIM59) used for forensic age prediction in three groups of individuals with diagnosed medical conditions. The obtained results showed that the selected age-related CpG sites have unchanged age prediction capacity in the group of late onset Alzheimer's disease patients. Aberrant hypermethylation and decreased prediction accuracy were found for TRIM59 and KLF14 markers in the group of early onset Alzheimer's disease suggesting accelerated aging of patients. In the Graves' disease patients, altered DNA methylation profile and modified age prediction accuracy were noted for TRIM59 and FHL2 with aberrant hypermethylation observed for the former and aberrant hypomethylation for the latter. Our work emphasizes high utility of the ELOVL2 and C1orf132 markers for prediction of chronological age in forensics by showing unchanged prediction accuracy in individuals affected by three diseases. The study also demonstrates that artificial neural networks could be a convenient alternative for the forensic predictive DNA analyses.

Snow sports related head and spinal injuries: an eight-year survey from the neurotrauma centre for the Snowy Mountains, Australia.

Neurotrauma from snow-sports related injuries is infrequently documented in the literature. In Australia no collective data has ever been published. The aim of this study is to document the injury pattern of snow sports related neurotrauma admissions to The Canberra Hospital, the regional trauma centre for the Snowy Mountains. A computerised hospital record search conducted between January 1994 and July 2002 revealed 25 head and 66 spinal injury admissions. The incidence of severe injuries requiring referral to tertiary trauma hospital was estimated to be 7.4 per 100,000 skier-days and for head and spinal injury 1.8 per 1,000,000 skier-days and 5.6 per 1,000,000 skier-days, respectively. Collision with a stationary object was disproportionately associated with head injury ( [Formula: see text] ) and falling forward with spinal injury ( [Formula: see text] ). Snowboarders tended to sustain cervical fractures more often than skiers ( [Formula: see text] ). The importance of helmet usage in buffering the impact of head-on collision and the proposition of having both feet fastened to a snowboard in leading to cervical injury were highlighted.

Angioplasty and stenting in the posterior cerebral circulation.

PURPOSE: To report initial experience with intracranial vertebral artery (VA) and basilar artery (BA) percutaneous transluminal angioplasty (PTA) and stenting. METHODS: Eleven patients (10 men; mean age 66 years, range 56-75) with intracranial 14 VA and 3 BA stenoses were managed from December 1997 through November 1999. All patients presented with vertebrobasilar ischemia (VBI) despite antiplatelet and anticoagulant therapy. Clinical presentations included visual disturbance (n = 8), dysarthria/dysphasia (n = 5), and vertigo (n = 5). RESULTS: Five patients underwent PTA only of 7 lesions with a mean preprocedural stenosis of 80% (range 50%-90%) that was reduced to 54% (range 30%-70%) after dilation. Six patients received 9 stents, 2 for VA dissections, 3 for tandem lesions, and 1 for a BA lesion. There were no embolic strokes. Patients were followed by clinical evaluation and personal or telephone interviews. Over a mean 18-month follow-up (range 12-35), 8 (73%) patients remained asymptomatic, while 3 (27%) had permanent deficits, 2 related to the procedure and 1 owing to distal disease CONCLUSIONS: Intracranial VA or BA angioplasty and stenting alleviated symptoms in patients with vertebrobasilar ischemia despite best medical management and may prevent stroke.

Microsurgery for small arteriovenous malformations of the brain: results in 110 consecutive patients.

OBJECTIVE: To examine the results of surgery in 110 consecutive patients with arteriovenous malformations (AVMs) smaller than 3 cm in diameter. These results are compared with the published results of other microsurgical series as well as with results for patients treated with focused irradiation. METHODS: From January 1989 to November 1998, 121 patients with AVMs smaller than 3 cm were treated at our institution. One hundred ten patients underwent microsurgical removal of their AVMs. The presentation, preoperative neurological status, and postoperative outcome were recorded. Follow-up was complete for all surgical cases. RESULTS: Of the 110 patients, 109 (99%) had angiographically confirmed obliteration of their AVMs. Two patients (1.8%) required reoperation for residual AVM. Two (4.3%) of 46 patients with AVMs in eloquent brain areas experienced worsening of their neurological status after surgery. One (1.6%) of 64 patients was worse neurologically after removal of an AVM in a noneloquent area. CONCLUSION: Microsurgical removal is a safe and effective treatment for the majority of AVMs smaller than 3 cm in diameter. Although the treatment is accompanied by a risk of acute onset of neurological deficits, this tends to be transient in the majority of cases. Furthermore, microsurgical excision of small AVMs offers patients immediate protection from the natural history of their vascular lesions.

Production of curdlan-type polysaccharide by Alcaligenes faecalis in batch and continuous culture.

The biosynthesis of a thermogelable, extracellular homo-beta-(1 leads to 3)-glucan called "curdlan," has been studied in batch and continuous cultures of Alcaligenes faecalis var. myxogenes. Curdlan production is associated with the poststationary phase of a nitrogen-depleted, aerobic batch culture. Exopolymer is not detected in single-stage, carbon-limited continuous cultures but curdlan can be isolated from the effluent of a nitrogen-limited chemostat operating at a dilution rate (D) of less than 0.1 h-1. A spontaneous variant of strain ATCC 21680 was isolated and found to be compatible with long-term, nitrogen-limited chemostat culture. The specific rate of curdlan production is approximately four times higher in poststationary batch cultures than in single-stage continuous fermentations. The product yield (Yp/S) associated with batch processing (nongrowing cultures) is approximately 0.5 g curdlan/g glucose, with CO2 being the only detectable by-product.

The use of bee venom melittin to assess the topography of membrane vesicles derived from Paracoccus denitrificans.

There exists considerable controversy regarding membrane topography in vesicles derived by osmotic lysis of spheroplasts of Gram-negative bacteria. It has been reported by others that bee venom can be used to quantitate the portion of a heterogeneous vesicle population with an inside-out orientation by determining the degree of loss of crypticity of NADH dehydrogenase activity. We have demonstrated that a major component of bee venom, melittin, causes an increase in the activity of several different respiratory enzymes in isolated membrane vesicles of Paracoccus denitrificans. The degree of stimulation produced by melittin is dependent upon (i) the nature of the respiratory substrates, (ii) the pH, (iii) the presence of Mg2+, (iv) the melittin: membrane protein ratio, and (v) the growth history of the cells from which the membrane vesicles were derived. Melittin-induced enhancement of TMPD:ascorbate and cytochrome c oxidase activities cannot be accounted for by increased accessibility of nonpermeant substrate to the interior of the vesicle. The stimulatory effect of melittin may rely in part on its ability to alter the proton permeability of the membrane thereby abolishing respiratory control. Collectively these observations call into question the usefulness of bee venom melittin in quantitative analyses of membrane topography. These results are consistent with the postulated existence of a homogeneous vesicle population in which the topography of the NADH dehydrogenase is different from that of the intact cell.

Physiology of Candida utilis yeast in zinc-limited chemostat culture.

Candida utilis NRRLY -900 was grown in aerobic continuous culture in a minimal salts medium with sucrose (1% w/v) as the carbon source. increasing the concentration of zinc in the medium from 2.3 microM results in an increase in the apparent critical dilution rate from 0.3 to 0.47h-1, and in the maximum biomass productivity form 1.5g dry weight per litre per hour (at D=0.33-1) to 2.56g per litre per hour (at D = 0.45-1). The maximum steady-state level of cell-associated zinc (at D = 0.4-1) is 4nmol Zn2+/mg dry weight, during carbon-limited growth, and about 9mumol Zn2+/mg dry weight when FeCI3 is omitted form the medium. At input zinc concentrations <20microM, the cellular zinc concentration decreases linearly in proportion to the input zinc concentration and at <4.5microM Zn2+ the culture becomes zinc-limited. Zinc-limitation results in a decrease iun the growth yield Ysurcosefrom 0.54 to 0.38 cells/g sucrose consumed, and Y0 from 22 to 13g cells/g-atom 02, suggesting an altered efficiency of energy metabolism. The composition of the culture biomass with respect to protein and RNA content is not affected by zinc limitation.

Hyperaccumulation of zinc by zinc-depleted Candida utilis grown in chemostat culture.

The steady-state levels of zinc in Candida utilis yeast grown in continuous culture under conditions of zinc limitations are <1nmol Zn2+/mg dry weight of cells. Unlike carbon-limited cells, zinc-depleted cells from a zinc-limited chemostat possess the capacity to accumulate and store zinc at levels far in excess of the steady-state level of 4 nmol/mg dry biomass observed in carbon-limited chemostat cultures. Zinc uptake is energy-dependent and apparently undirectional since accumulated 65Zn neither exists from preloaded cells nor exchanges with cold Zn2+. The transport system exhibits a high affinity for Zn2+ (Km =.36micrM) with a Vmaxof 2.2 nmol per minute per milligram dry weight of cells. Growth during the period of the uptake assay is responsible for the apparent plateau level of 35 nmol Zn2+/mg dry weight of cells achieved after 20-30 min in the presence of 65Zn at pH 4.5 and 30 degrees C. Inhibition of growth during the uptake assay by cycloheximide results in a biphasic linear pattern of zinc accumulation where the cellular zinc is about 60 nmol/mg dry weight after 1 h. The enhanced level of accumulated zinc is not inhibtory to growth. Zinc-depleted C. utilis contains elevated amounts of polyphosphate and this anionic evidence does not allow discrimination between possible regulation of zinc homestasis either by inhibitions of zinc efflux through control of the membrane carrier or by control of the synthesis of a cytoplasmic zinc-sequestering macromolecule.

Subcellular distribution of lysophosphatidylinositol and lysophosphatidylcholine acyltransferases in rat brain.

Lysophosphatidylinositol acyltransferase utilizing arachidonoyl CoA and lysophosphatidylcholine acyltransferase utilizing linoleoyl CoA were measured in subcellular fractions of rat brain. In general, the distribution of activities paralleled that of NADPH--cytochrome c reductase. It is concluded that the endoplasmic reticulum is the major site of these acyltransferase activities in rat brain.