Predictive performance of Acute Physiological and Chronic Health Evaluation releases II to IV: a single New Zealand centre experience.

There is debate in Australia and New Zealand around the appropriate use of illness severity scoring systems in Australasian intensive care units. The international benchmark is the Acute Physiological and Chronic Health Evaluation (APACHE) system. In order to compare the performance of recent APACHE releases, we audited 2080 sequential patients admitted between 1 January 2006 and 31 March 2008 to the Middlemore Hospital intensive care unit, Auckland, New Zealand. We compared the predictive performance of the proprietary APACHE II, IIIh, IIIj and IV releases, and the performance of a 'localised' version of APACHE II containing re-estimated coefficients derived from a legacy dataset (7703 sequential patients admitted between 1 January 1997 and 31 December 2005). Discrimination assessed by receiver operating characteristic curves was highest with the APACHE III and IV releases, and significantly better than the APACHE II releases. Calibration assessed by the Hosmer-Lemeshow statistic was poor with all releases, although it was best with APACHE IV and 'localised' version of the APACHE II release. Overall accuracy assessed by the Brier Mean Probability score and Shapiro's R statistic was best with APACHE IV. Our study suggests the possibility of improved prediction in moving to APACHE IV from older releases, although broader multicentre study within the Australian and New Zealand critical care community is warranted. Our study also suggests localisation of the APACHE system offers further opportunity to improve prediction, although these improvements may not be major without ground-up development of a new risk prediction model within our local critical care setting.

Histopathologic and physiologic effects of chronic implantation of microelectrodes in sacral spinal cord of the cat.

Active microelectrodes were implanted for a period of 2 weeks to 3 months into the sacral spinal cord of 10 male cats in order to test the feasibility and the safety of discrete stimulation of the parasympathetic preganglionic nucleus for future clinical applications of microelectrode technology in micturition control. An array of four 50 microns-diameter iridium microelectrodes was inserted beneath the dura in each cat. At weekly intervals, bladder pressure was measured as hydrostatic pressure on an intraluminal catheter. At the end of the period, histopathology was evaluated with serial transverse epoxy sections. Observations included diffuse and focal axonal degeneration in white matter and possible neuronal loss around the electrode in the gray matter, meningeal ensheathment of the shafts, and occasional aseptic inflammation of tissue and apparent movement of the electrodes after implantation. Increased bladder pressure responses to individually pulsed electrodes located within the sacral parasympathetic nucleus were not consistent, and, surprisingly, at least 2 different sites were also effective. As long as 3 months after implantation, in 2 out of 5 animals, pulsing of electrodes consistently produced micturition. We conclude that while microelectrode implants are feasible, further modifications in electrode design are needed to eliminate movement and inflammation.

Magnetite biomineralization in the human brain.

Although the mineral magnetite (Fe3O4) is precipitated biochemically by bacteria, protists, and a variety of animals, it has not been documented previously in human tissue. Using an ultrasensitive superconducting magnetometer in a clean-lab environment, we have detected the presence of ferromagnetic material in a variety of tissues from the human brain. Magnetic particle extracts from solubilized brain tissues examined with high-resolution transmission electron microscopy, electron diffraction, and elemental analyses identify minerals in the magnetite-maghemite family, with many of the crystal morphologies and structures resembling strongly those precipitated by magnetotactic bacteria and fish. These magnetic and high-resolution transmission electron microscopy measurements imply the presence of a minimum of 5 million single-domain crystals per gram for most tissues in the brain and greater than 100 million crystals per gram for pia and dura. Magnetic property data indicate the crystals are in clumps of between 50 and 100 particles. Biogenic magnetite in the human brain may account for high-field saturation effects observed in the T1 and T2 values of magnetic resonance imaging and, perhaps, for a variety of biological effects of low-frequency magnetic fields.

Alteration of the Magnetic Properties of Aquaspirillum magnetotacticum by a Pulse Magnetization Technique.

The presence of a narrow shape and size distribution for magnetite crystals within magnetotactic organisms suggests strongly that there are species-specific mechanisms that control the process of biomineralization. In order to explore the extent of this control, cultures of Aquaspirillum magnetotacticum in the exponential growth phase were exposed to increasing magnetic pulses with the aim of separating cell populations on the basis of their magnetic coercivities. Isothermal remanent magnetization and anhysteretic remanent magnetization studies were performed with freeze-dried magnetic cells after the remagnetization treatment. Subpopulations of A. magnetotacticum that showed an increase in coercivity correlated with the intensity of the magnetic pulses were isolated. After successive subcultures of the remaining north-seeking cells, a maximum bulk coercivity (H(b)) of 40 mT was obtained after treatment with a 55-mT pulse. Although we obtained A. magnetotacticum variants displaying higher coercivities than the wild-type strain, changes in crystal size or shape of the magnetite crystals were below reliable detection limits with transmission electron microscopy. Attempts to shift the coercivity towards higher values caused it to decrease, a change which was accompanied by an increase in magnetostatic interactions of the magnetosome chains as well as an increase in the cell population displaying an abnormal distribution of the magnetosome chains. Ultrastructural analyses of cells and magnetosomes revealed the appearance of cystlike bodies which occasionally contained magnetosomes. The increase in cystlike cells and abnormal magnetosome chains when higher magnetic pulses were used suggested that magnetosomes were collapsing because of stronger interparticle magnetostatic forces.

Uptake of tritiated thymidine in mitochondria of the retina.

Light microscopic autoradiography with 3H-thymidine was used as a probe for DNA synthesis on the retinas of the guinea pig, rabbit, and monkey. Relatively heavy labeling was found in the ganglion cell and inner nuclear layers as well as in the photoreceptor inner segments in the guinea pig and monkey. Electron microscopic autoradiography demonstrated that in the ganglion cells and photoreceptor inner segments in the monkey, 89% of silver grains, representing thymidine uptake, were on or near the mitochondria.

Localization of actin and tubulin in developing and adult mammalian photoreceptors.

In order to define cytoskeletal domains of the mammalian photoreceptor, actin and tubulin were localized in adult retinae of mouse and human. For light-microscopic localization, actin was labeled using fluorescent phalloidin or monoclonal antibodies against actin, and tubulin was labeled using monoclonal antibodies against alpha- and beta-tubulin in an immunocytochemical method. Actin and tubulin were also localized by ultrastructural immunocytochemistry in the mouse. Filamentous actin was present in the retina at the outer limiting membrane and in synaptic terminals, especially of the cones, while globular actin was observed additionally in the inner segments. Müller cell cytoplasm and apical microvilli at the outer limiting membrane were also labeled for filamentous actin. Alpha- and beta-tubulin were evident throughout the photoreceptors, including the inner segments, but not in the synaptic terminals or at the outer limiting membrane. In the early postnatal retina of mouse, actin and tubulin were present at the ventricular surface. This pattern changed as photoreceptors fully elongated and as synaptogenesis occurred in the outer plexiform layer.

Distribution of ascorbate in normal primate retina and after photic injury: a biochemical, morphological correlated study.

The levels of reduced and oxidized ascorbates were determined in the normal baboon neural retina and pigment epithelium-choroid. The ascorbate in the neural retina was mainly in the reduced form, while in the pigment epithelium, it existed primarily in the oxidized form. One eye of each of six baboons was exposed to indirect ophthalmoscope light for 30 minutes. Morphologic study showed necrotic photoreceptors, outer segment disorganization, and abnormal pigment epithelial basal infoldings and swollen microvilli. After the light exposure, the values of the reduced ascorbate decreased in both the neural retina and pigment epithelium-choroid of the posterior pole. The possible role of ascorbate as an antioxidant and ascorbate transport into the retina are discussed.

Effect of photic injury on the retinal tissues.

In order to study the tissue damages in photic injury of the retina, the macula and posterior pole of the rhesus monkey were exposed to the light of an indirect ophthalmoscope. During the acute phase of the injury, disruption of the blood-retinal barrier appeared to be an early and sensitive indicator of retinal pigment epithelial damage and cyclic-GMP-phosphodiesterase of the photoreceptor lamella was inactivated. These lesions were followed for 2 to 5 years. Focal chronic decompensation of the blood-retinal barrier and subretinal pigment epithelial neovascularization were noted in the chronic degenerative phase of this photic maculopathy. A possible protective role of ascorbic acid in mild photic injury was proposed.

Reduced and oxidized ascorbates in guinea pig retina under normal and light-exposed conditions.

Both reduced and oxidized ascorbates were measured in aqueous, neural retina, and pigment epithelium-choroid complex (PE-C) of pigmented guinea pigs. Normal values for total ascorbate of 16 mg/dl in aqueous, 22 mg/dl in neural retina, and 7 mg/dl in PE-C were found. After mild photic damage caused by varying lengths of exposure of 10,000 to 20,000 lux of fluorescent lighting, reduced ascorbate concentrations generally decreased in the neural retina, while oxidized ascorbate generally increased in PE-C. In both normal and light-exposed retinas, reduced ascorbate was predominant in the neural retina, and oxidized ascorbate was predominant in the PE-C. Histochemical localization of reduced ascorbate occurred in the Müller cell fibers and at the apices of the retinal pigment epithelium.