Validation of the Applied Biosystems Prism 377 automated sequencer for the forensic short tandem repeat analysis

The Applied Biosystems (ABI) Prism 377 DNA sequencer has been evaluated in an attempt to increase the throughput of samples for short tandem repeat (STR) analysis, in both forensic casework and the UK National Criminal Intelligence DNA Database. The gel system assessed consisted of 0.2 mm, 4% acrylamide 6 M urea gels, with a well-to-read distance of 36 cm. Gels were run at a constant voltage of 3 kV and constant temperature of 51 degrees C. The run time of our second generation multiplex (SGM) STR system was achieved in less than 2 h. Rigorous validation has been performed on the instrument hardware and software. Complete resolution of 1 base differences was obtained, up to and beyond 350 bases; sizing precision across gels was more than 2-fold higher than the 373A and the sensitivity was increased by one third.     

The effect of unilateral destruction of the head of the caudate nucleus with kainic acid on the structure of the wakefulness-sleep cycle and the EEG in rats

The wakefulness-sleep cycle characteristics changes in Wistar line rats after unilateral lesions of the neuronal elements of the caudate nucleus head with kainic acid (at sparing the conducting ways going through this structure) were studied. It was shown that in 4 days after acid injection the share of the active wakefulness and light slow-wave sleep is significantly increased, whereas the percentage of the passive wakefulness, deep slow-wave stage, and paradoxal sleep (up to 80%) is reduced. The wakefulness-sleep cycle in rats is completely normalized on the 7th day after injection. The obtained data allow to consider the caudate nucleus as a modulator for wakefulness-sleep cycle, that is the structure involved in the motive activity regulation.     

Effect of intrathecal morphine on blood glucose, glucagon and tissue glycogen in rat: comparison with the effect of xanthan gum on blood glucose

Eleven patients suffering from chronic disabling tinnitus underwent an FDG-PET study (positron emission tomography with [18F]deoxyglucose). Nine tinnitus patients revealed a significantly increased metabolic activity in the left, 1 in the right primary auditory cortex (PAC, Brodmann area 41). These results were statistically significant when compared to 14 healthy control individuals without tinnitus. A negative result was obtained from a chronic tinnitus patient but who had no subjective complaints during the period of PET investigation. One patient was first investigated during a disabling tinnitus period, later during a period with tinnitus relief and again when suffering from severe tinnitus. The metabolic activity of his left PAC was in good accordance with the subjective degree of tinnitus complaints present during each PET investigation. Although for the first time these results give objective evidence of tinnitus sensation and localization, they are difficult to interpret because of the limited research data available that combine functional brain imaging and acoustic stimuli.     

Morphometric video analysis of the engine-driven nickel-titanium Lightspeed instrument system

Two hundred and sixteen Lightspeed instruments were evaluated microscopically for the presence of corrosion, surface debris, and alloy defects. The instruments were assessed morphometrically for consistency of physical design and dimensions by measuring and analyzing eight parameters of the instrument pilot tips, heads, and shafts. Results from visual inspection showed that none of the instruments were corroded; 23 presented surface porosities, and 17 had sharp strips of alloy. Data obtained by morphometric analysis indicated the mean diameter of the head of only 7 of 18 sizes met the +/- 0.02 mm allowable tolerance set forth by the American Dental Association (ADA) Specification No. 28. Observation and video analysis indicated that instruments of the same size adhere to the same basic design, but that morphometric variations do exist. The visual and intersize analysis indicated that the Lightspeed is not an instrument of any one determined shape that changes only in diameter. Rather, it is a series of instruments that show gradual shifts in both size and shape as the instrument size increases. Lightspeed instruments are a new type of nickel-titanium endodontic instrument that cannot be evaluated using the standards proposed by the American National Standards Institute/ADA Specification No. 28 for files and reamers.     

Identification of a novel inhibitor (NSC 665564) of dihydroorotate dehydrogenase with a potency equivalent to brequinar

A novel inhibitor of dihydroorotate dehydrogenase (DHO-DH) has been discovered using data from the National Cancer Institute's in vitro drug screen. Upon analysis of cytotoxicity results from the sixty tumor cell lines used in this screen, the COMPARE program predicted that NSC 665564 was likely to have the same mechanism of inhibition as brequinar, a known potent inhibitor of DHO-DH. We validated this prediction experimentally using MOLT-4 lymphoblast and found the IC50 of brequinar (0.5 microM) and NSC 665564 (0.3 microM) were comparable and that this induced cytotoxicity was reversed by either uridine or cytidine. The enzyme target of NSC 665564 was shown to be identical to that of brequinar when incubation with each drug followed by a 1 h pulse with [14C] sodium bicarbonate resulted in cellular accumulation of [14C]N-carbamyl-L-aspartic acid and [14C]L-dihydroorotic acid, with concurrent marked depletion of CTP and UTP. The Ki's for NSC 665564 and brequinar were 0.14 and 0.24 microM, respectively, when partially purified MOLT-4 mitochondria (the site of DHO-DH) were used. These results show that mechanistic predictions obtained using correlations from the COMPARE algorithm are independent of structure since the structure of NSC 665564 is dissimilar to that of other established DHO-DH inhibitors.     

Ambulatory urodynamics: extramural testing of the lower and upper urinary tract by Holter monitoring of cystometrogram, uroflowmetry, and renal pelvic pressures

This article elucidates the clinical applicability and state of the art of ambulatory urodynamics. Ambulatory urodynamics have evolved into practical investigations like EAC, HFM, and EAC combined with renal pelvimetry. EAC has been shown to be the method of preference if detrusor overactivity is involved. Conventional filling cystometry has proved to be an unreliable way to exclude detrusor instability. De novo instability after suspension surgery often indicates that an existing detrusor overactivity was not identified preoperatively. EAC including flowmetry has shown considerable variance in obstructive and contractility parameters in males with LUTS indicative for BPH. This raises doubt whether the clinical flow analysis is the suitable "gold standard" as advocated by the ICS. For a real break through of EAC, less complex automatic analysis is necessary. HFM is a newer method within the range of ambulatory urodynamic tests. It has not yet been completely evaluated. But, because the technique is analogous to the office flowmetry, noninvasive and very well accepted by the patients, it is expected to be widely used. This expectation is strengthened by the fact that HFM seems to show individual therapeutic efficacy of drugs, such as alpha-blockers. As a research tool to evaluate efficacy, it is far more powerful than conventional methods because of the reduction of within-patient standard deviation to about 10%. Finally, EAC combined with pelvimetry offers a promising method for the clinical evaluation of a combined dysfunction of upper and lower urinary tract.     

A gene which causes severe ocular alterations and occipital encephalocele (Knobloch syndrome) is mapped to 21q22.3

Knobloch syndrome (KS), characterized by high myopia, vitreoretinal degeneration with retinal detachment, macular abnormalities and occipital encephalocele, was recently confirmed as autosomal recessive. Here we report the assignment of the gene for this syndrome to 21q22.3 with the marker D21S171 through homozygosity mapping in a highly inbred Brazilian family with 11 affected individuals. A total of nine markers spanning a region of 15.2 cM of the chromosome 21q22.3 were tested and the candidate region was restricted to an interval of 4.3 cM.     

Prosthetic valve endocarditis: superiority of surgical valve replacement versus medical therapy only

The objective of our study was to assess the long-term outcome of patients with prosthetic valve endocarditis. We used a multicenter, prospective, observational study design. Six university teaching hospitals with high volume cardiothoracic surgery participated. Seventy-four patients with prosthetic valve endocarditis as defined by explicit, objective criteria were selected for participation. All patients were followed up prospectively for 1 year. Thirty-one percent and 69% had development of endocarditis within 60 days of valve insertion ("early") and after 60 days ("late"), respectively. The most common causes were Staphylococcus epidermidis (40%), Staphylococcus aureus (20%), streptococcal species (18%), and aerobic gram-negative bacilli (11%). Physical signs of endocarditis (new or changing murmur, stigmata, emboli) were seen in 58%. At 6 months and 12 months, mortality was 46% and 47%, respectively. Surgical replacement of the infected valve led to significantly lower mortality (23%) as compared with medical therapy alone (56%), as assessed by both univariate and multivariate analyses (p < 0.05). Improved outcome was seen for the surgical group even when controlling for severity of illness at time of diagnosis. From these findings we conclude that accurate assessment of outcome in prosthetic valve endocarditis requires long-term follow-up of at least 6 months following diagnosis. Surgical therapy warrants greater scrutiny; evaluation in controlled clinical trials is appropriate.