Multiplexed DNA sequencing and diagnostics by hybridization with enriched stable isotope labels

A new DNA diagnostic and sequencing system has been developed that uses time-of-flight resonance ionization mass spectrometry (TOF-RIMS) to provide a rapid method of analyzing stable isotope-labeled oligonucleotides in form 1 sequencing by hybridization (SBH). With form 1, the DNA is immobilized on a nylon membrane and enriched isotope-labeled individual oligonucleotide probes are free to seek out complementary DNAs during hybridization. The major advantage of this new approach is that multiple oligonucleotides can be labeled with different enriched isotopes and can all be simultaneously hybridized to the genosensor matrix. The probes can then be simultaneously detected with TOF-RIMS with high selectivity, sensitivity, and efficiency. By using isotopically enriched tin labels, up to 10 labeled oligonucleotides could be examined in a single hybridization to the DNA matrix. Greater numbers of labels are available if rare earth isotopes are employed. In the present study, matrices containing three different DNAs were prepared and simultaneously hybridized with two different probes under a variety of conditions. The results show that DNAs, immobilized on nylon surfaces, can be specifically hybridized to probes labeled with different enriched in isotopes. Discrimination between complementary and noncomplementary sites of better than 100 was obtained in multiplexed samples. This new SBH method, which employs stable isotopic labels to locate target DNAs and TOF-RIMS to detect the labels, will be a very versatile and extensive multiplexing method.     

Age- and sex-related differences in nuclear lipid content and nucleoside triphosphatase activity in the JCR:LA-cp corpulent rat

The putative role of the nuclear nucleoside triphosphatase (NTPase) is to provide energy to the nuclear pore complex for poly A(+) mRNA export. Previous work has demonstrated that liver nuclear NTPase activity is greater in 6 month old corpulent (cp/cp) female JCR:LA rats, a hyperlipidemic rat model, compared to lean (+/?) animals. This increase appeared to be related to increases in nuclear membrane cholesterol content. The current study extended these initial data to compare NTPase activity as a function of age and sex in isolated JCR:LA-cp rat liver nuclei, to further test the hypothesis that nuclear membrane cholesterol may modulate NTPase activity. NTPase activity was increased in cp/cp female animals compared to +/? females at all ages studied, with Vmax values increased by 60-176%. Membrane integrity of cp/cp female nuclei was reduced compared to +/? female nuclei. Nuclear membrane cholesterol levels increased linearly with age by 50, 150 and 250% in 3, 6 and 9 month old cp/cp females over leans. In contrast, nuclei from cp/cp males exhibited only minor, isolated changes in NTPase activity. Furthermore, there were no significant changes in nuclear cholesterol content or membrane integrity in the less hyperlipidemic male animals at any age. These data suggest that altered lipid metabolism may lead to changes in nuclear membrane structure, which in turn may alter NTPase activity and functioning of the nuclear pore complex.     

Hepatic hemangioma. Analysis of 103 cases. Clinical considerations and imaging methods

Hemangiomas are the most common benign vascular tumor found in the liver. In this study, 103 cases of hepatic hemangiomas were analysed in a retrospective manner. Image aspects such as those seen in ultrassonography, computed tomography, nuclear magnetic resonance and arteriography are discussed. The incidence was higher amongst the female sex, between individuals from 40 to 60 years old, which presented predominantly without symptoms (66%) and localized mainly in the right hepatic lobe (74%). The meaning of those aspects, when adopted, permits the clinicians to differentiate benign from malignant nodular lesions in the liver.     

The vestibulo-collic reflex

This study was undertaken to compare the fixation of the semitubular blade plate with that of the AO T plate. Cadaveric humeri (n=12 pairs) from an elderly population (41 to 89 years) had either a blade plate (n=12) or a T plate (n=12) fixed to them, subsequent to which a transverse osteotomy was performed. Mechanical testing in tension was performed in 1 series (n=5, blade plate; n=5, T plate) by applying a single maximal load to failure and in a second series (n=7, blade plate; n=7, T plate) by applying submaximal cyclic loading before failure was induced. Results showed that the fixation provided by the semitubular blade plate was significantly better (P < .05) than that of the T plate in those specimens subjected to submaximal cyclic loading before failure (series 2). This latter testing method contains some component of the clinical situation compared with monotonic distraction to failure. Based on these results plus favorable clinical results reported in the literature, the semitubular blade plate is possibly a better alternative to the T plate in the management of proximal humeral fractures that require open reduction and internal fixation.     

Spray‐Deposited Al‐Si (Osprey CE) Alloys and their Properties

Osprey Metals Ltd. has developed a new family of lightweight, low expansion aluminium‐silicon alloys to suit electronic applications, containing between 30 and 70 wt.% aluminium. They are produced using the Osprey spray deposition process, which achieves homogeneous and isotropic properties. Their advantageous physical characteristics, combining low thermal expansion, high thermal conductivity and low density are particularly suited for packaging electronic circuitry. These CE (controlled expansion) alloys are easy to machine to tight tolerances using standard machine tools and they can be electroplated with gold, silver or nickel finishes without difficulty. This paper focuses on the measurement of mechanical (flexural strength) and thermo‐mechanical properties (CTE, thermal diffusivity and specific heat) of these alloys, carried out in collaboration with AMTT. The property measurements were consistent with those measured previously elsewhere, apart from the CTE values, which deviated significantly and were at variance with values predicted from a rule of mixtures. This discrepancy was resolved in a short follow‐up series of measurements carried out under conditions corresponding more closely to temperature equilibrium.