Potentiometric determination of coenzyme-dependent oxidoreductase enzymes with recycling of the coenzyme

By enzymatically establishing a rapid (essentially equilibrium) coupling of a redox coenzyme such as NAD with the components of the ferrocyanide–ferricyanide half-cell (e.g., using excess diaphorase) the half-cell potential can be used to monitor another enzymatic reaction involving the same coenzyme. This approach provides a general, rapid potentiometric method of assaying coenzyme-dependent oxidoreductase enzymes. We show that these assay systems can be designed for multiple turnover of coenzyme (in our case NAD) during a single assay thereby amplifying the rate of electromotive force (emf) change with a concomitant increase in sensitivity of enzyme assay. This allows the use of small concentrations of coenzyme and extension of the range of enzyme concentrations that may be assayed.     

Morphological and histochemical characterization of the pectoral fin muscle of batoids

Batoids differ from other elasmobranch fishes in that they possess dorsoventrally flattened bodies with enlarged muscled pectoral fins. Most batoids also swim using either of two modes of locomotion: undulation or oscillation of the pectoral fins. In other elasmobranchs (e.g., sharks), the main locomotory muscle is located in the axial myotome; in contrast, the main locomotory muscle in batoids is found in the enlarged pectoral fins. The pectoral fin muscles of sharks have a simple structure, confined to the base of the fin; however, little to no data are available on the more complex musculature within the pectoral fins of batoids. Understanding the types of fibers and their arrangement within the pectoral fins may elucidate how batoid fishes are able to utilize such unique swimming modes. In the present study, histochemical methods including succinate dehydrogenase (SDH) and immunofluoresence were used to determine the different fiber types comprising these muscles in three batoid species: Atlantic stingray (Dasyatis sabina), ocellate river stingray (Potamotrygon motoro) and cownose ray (Rhinoptera bonasus). All three species had muscles comprised of two muscle fiber types (slow-red and fast-white). The undulatory species, D. sabina and P. motoro, had a larger proportion of fast-white muscle fibers compared to the oscillatory species, R. bonasus. The muscle fiber sizes were similar between each species, though generally smaller compared to the axial musculature in other elasmobranch fishes. These results suggest that batoid locomotion can be distinguished using muscle fiber type proportions. Undulatory species are more benthic with fast-white fibers allowing them to contract their muscles quickly, as a possible means of escape from potential predators. Oscillatory species are pelagic and are known to migrate long distances with muscles using slow-red fibers to aid in sustained swimming.     

Human Cytomegalovirus UL50 and UL53 Recruit Viral Protein Kinase UL97, Not Protein Kinase C,for Disruption of Nuclear Lamina and Nuclear Egress in Infected Cells

Herpesvirus nucleocapsids traverse the nuclear envelope into the cytoplasm in a process called nuclear egress that includes disruption of the nuclear lamina. In several herpesviruses, a key player in nuclear egress is a complex of two proteins, whose homologs in human cytomegalovirus (HCMV) are UL50 and UL53. However, their roles in nuclear egress during HCMV infection have not been shown. Based largely on transfection studies, UL50 and UL53 have been proposed to facilitate disruption of the nuclear lamina by recruiting cellular protein kinase C (PKC), as occurs with certain other herpesviruses, and/or the viral protein kinase UL97 to phosphorylate lamins. To investigate these issues during HCMV infection, we generated viral mutants null for UL50 or UL53. Correlative light electron microscopic analysis of null mutant-infected cells showed the presence of intranuclear nucleocapsids and the absence of cytoplasmic nucleocapsids. Confocal immunofluorescence microscopy revealed that UL50 and UL53 are required for disruption of the nuclear lamina. A subpopulation of UL97 colocalized with the nuclear rim, and this was dependent on UL50 and, to a lesser extent, UL53. However, PKC was not recruited to the nuclear rim, and its localization was not affected by the absence of UL50 or UL53. Immunoprecipitation from cells infected with HCMV expressing tagged UL53 detected UL97 but not PKC. In summary, HCMV UL50 and UL53 are required for nuclear egress and disruption of nuclear lamina during HCMV infection, and they recruit UL97, not PKC, for these processes. Thus, despite the strong conservation of herpesvirus nuclear egress complexes, a key function can differ among them.     

Iridium-192 brachytherapy in combination with interstitial microwave-induced hyperthermia for malignant glioma

A phase I clinical trial assessing the feasibility and safety of hyperthermia in combination with 192Ir brachytherapy (60 Gy) for the treatment of malignant glioma now includes 14 patients. Hyperthermia (42-43 degrees C at tumor margin for 60 min) has been induced using stereotactically implanted afterloading catheters and a microwave (915 or 2,450 MHz) antenna array. Thermometry recorded along each catheter confirms the general ability of the technique to heat such volumes, but thermal heterogeneities are documented. Transient or permanent worsening of previous neurologic deficit, seen in 7 patients, has been the most common morbidity.     

Predator prey relationships under luminous conditions

Lemon sharks seized lighted grey snappers before seizing the unlighted but otherwise identical grey snappers in ten consecutive observations. Snappers bearing a light embedded in epoxy resin were approached in a straight line attack pattern while (dark) controls were always circled by the shark at least once before attack and seizure.     

Preparation and properties of soluble-insoluble nicotinamide coenzymes.

A soluble-insoluble form of nicotinamide adenine dinucleotide (NAD+), which can be rendered either soluble or insoluble by simply adjusting the pH, has been prepared by covalently coupling NAD to alginic acid using 1,2,7,8-diepoxyoctane. The NAD bound to the alginic acid showed the coenzymic function in the soluble state and could be collected for further use as precipitate by lowering the pH to below 3. Coupling soluble-insoluble coenzymes with insolubilized apoenzymes is possible in fluidized and fixed-bed reactors.     

Pre location by clownfish (Amphiprion perideraion) larvae feeding on rotifers (Brachionus plicatilis)

Prey location by clownfish (Amphiprion perideraion) larvae feedingon rotifers was accurately characterized. Cruise predator cloworishlarvae locate prey throughout a threedimensional search space,not only on the boundaries of this space as previously predicted.The search space ranged from 0 to 120° away from the lengthaxis of the fish in the horizontal plane and from 0 to 150°in the transverse plane. The boundaries of the search spacewere 1.3 body lengths away along the axis of the fish and 1body length away to the sides and above. First-feeding clownfishlarvae have well-developed visual acuity and binocular vision.