International year of Chemistry 2011. A guide to the history of clinical chemistry.

BACKGROUND: This review was written as part of the celebration of the International Year of Chemistry 2011. CONTENT: In this review we provide a chronicle of the history of clinical chemistry, with a focus on North America. We outline major methodological advances and trace the development of professional societies and journals dedicated to clinical chemistry. This review also serves as a guide to reference materials for those interested in the history of clinical chemistry. The various resources available, in sound recordings, videos, moving images, image and document archives, museums, and websites dedicated to diagnostic company timelines, are surveyed. SUMMARY: These resources provide a map of how the medical subspecialty of clinical chemistry arrived at its present state. This information will undoubtedly help visionaries to determine in which direction clinical chemistry will move in the future.

Mechanisms of aluminum-induced neurodegeneration in animals: Implications for Alzheimer's disease.

For four decades the controversial question concerning a possible role for aluminum neurotoxicity in contributing to the pathogenesis of Alzheimer's disease has been debated, and studies by different investigators have yielded contradictory results. The lack of sensitivity to aluminum neurotoxicity in transgenic mouse models of Alzheimer's disease has not allowed the system to be used to explore important aspects of this toxicity. Rabbits are particularly sensitive to aluminum neurotoxicity and they develop severe neurological changes that are dependent on dose, age and route of administration. The most prominent feature induced by aluminum in rabbit brain is a neurofibrillary degeneration that shares some similarity with the neurofibrillary tangles found in Alzheimer's disease patients. In the present review we discuss data from our laboratory and others, on the effects of aluminum on behaviour, neurologic function and morphology, using aluminum administered to rabbits via different routes. Finally, we will examine data on the possible cellular mechanisms underlying aluminum neurotoxicity, and potential neuroprotective strategies against aluminum toxicity.

Peri-nuclear clustering of mitochondria is triggered during aluminum maltolate induced apoptosis.

Synapse loss and neuronal death are key features of Alzheimer's disease pathology. Disrupted axonal transport of mitochondria is a potential mechanism that could contribute to both. As the major producer of ATP in the cell, transport of mitochondria to the synapse is required for synapse maintenance. However, mitochondria also play an important role in the regulation of apoptosis. Investigation of aluminum (Al) maltolate induced apoptosis in human NT2 cells led us to explore the relationship between apoptosis related changes and the disruption of mitochondrial transport. Similar to that observed with tau over expression, NT2 cells exhibit peri-nuclear clustering of mitochondria following treatment with Al maltolate. Neuritic processes largely lacked mitochondria, except in axonal swellings. Similar, but more rapid results were observed following staurosporine administration, indicating that the clustering effect was not specific to Al maltolate. Organelle clustering and transport disruption preceded apoptosis. Incubation with the caspase inhibitor zVAD-FMK effectively blocked apoptosis, however failed to prevent organelle clustering. Thus, transport disruption is associated with the initiation, but not necessarily the completion of apoptosis. These results, together with observed transport defects and apoptosis related changes in Alzheimer disease brain suggest that mitochondrial transport disruption may play a significant role in synapse loss and thus the pathogenesis or Alzheimer's disease.

Evaluation of analytical methods and workflow performance of the Architect ci8200 integrated serum/plasma analyzer system.

BACKGROUND: The Architect ci8200 is an integrated serum analyzer for photometric, electrochemical and immunological assays. Several assays of each category and the workflow performance of the system were compared with established laboratory procedures in two laboratories. METHODS: Measurements were compared with the ELECSYS 2010 (Roche Diagnostics) for CEA, PSA, FPSA, AFP, folate, vitamin B12, with the CENTAUR (Bayer) for TSH, T4, FT4, FSH and Estradiol, with the LIAISON (DiaSorin) for TSH, FT4 and FT3, with the Behring Nephelometer BN II (Dade-Behring) for ferritin, and with the INTEGRA 800 (Roche Diagnostics), and the AU640 (Olympus) for clinical chemistry assays. Workflow studies were performed to compare times of analysis required for defined analytical workloads. RESULTS: The coefficients of variation (CVs) for within-run imprecision were between 3% and 6% for CEA, PSA, FPSA, AFP and ferritin, and between 3% and 11% for TSH, FT4, FT3, folate and vitamin B12. The CVs for day-to-day imprecision for immunoassays were between 3% and 10%, except for vitamin B12 (CVs 11-13%) and FT4 (CV 10% -13%). For clinical chemistry tests corresponding CVs for within-run imprecision were < 1%, except for HDL, triglyceride, creatinine, ALT, LD and lipase (CVs<2%) and bicarbonate (CV 3%-6%) and magnesium (CV < 3%). The CVs for day-to-day imprecision for clinical chemistry tests were < 1%, except for sodium, CO(2), magnesium, phosphorus, glucose, uric acid, HDL, triglyceride, ALT, AST CK, lipase with CVs < 6% and for CO(2)<11%. Dilutional linearity testing of seven immunoassays and five clinical chemistry analytes resulted in recovery rates of 90-110%. Correlation studies with 15 immunoassays and 25 clinical chemistry tests showed acceptable agreements with established methods. Work flow analyses demonstrated a net gain in time of analysis up to 109 min depending on the size of the sample batch analyzed with the Architect ci8200 as the main analyzer as compared to the currently installed routine laboratory equipment. Median turn-around times were 7 and 30 min for chemistry assays and immunoassays, respectively, when ordered as STAT analyses, and 18 min when chemistry assays were ordered as routine determinations. CONCLUSIONS: Assays on the Architect ci8200 performed well, fulfilling quality control requirements as defined for instance by German quality control guidelines (RiliBAK). Method comparisons showed acceptable agreements with established assays. Workflow studies using the Architect ci8200 documented shorter times of analyses as compared with the conventionally established laboratory routine demonstrating the potential of integrated chemistry/immunoassay analyzers to provide faster and more efficient performance.

Aluminum maltolate-induced toxicity in NT2 cells occurs through apoptosis and includes cytochrome c release.

Aluminum (Al) compounds are neurotoxic and have been shown to induce experimental neurodegeneration although the mechanism of this effect is unclear. In order to study this neurotoxic effect of Al, we have developed an in vitro model system using Al maltolate and human NT2 cells. Al maltolate at 500 microM caused significant cell death with a 24-h incubation and this toxicity was even more evident after 48 h. Lower doses of Al maltolate were also effective, but required a longer incubation for cell death. Nuclear fragmentation suggestive of apoptosis was observed as early as three hours and increased substantially through 24 h. Chromatin condensation and nuclear fragmentation were confirmed by electron microscopy. In addition, TUNEL positive nuclei were also observed. The release of cytochrome c was demonstrated with Western blot analysis. This in vitro model using human cells adds to our understanding of Al neurotoxicity and could provide insight into the neurodegenerative processes in human disease.

GDNF regulates the A beta-induced endoplasmic reticulum stress response in rabbit hippocampus by inhibiting the activation of gadd 153 and the JNK and ERK kinases.

Glial cell line-derived neurotrophic factor (GDNF) is a potent survival agent for neurons, however, its effect on A beta-evoked neuronal death has not been examined. We show that the injection of A beta into New Zealand white rabbit brain activates the endoplasmic reticulum (ER) chaperones, grp 78 and grp 94, and the transcription factor, gadd 153. These effects correlate with the activation of JNK and ERK as well as of microglia and with the phosphorylation of tau protein. Treatment with GDNF inhibits the activation of gadd 153, reduces the phosphorylation of JNK, abolishes the phosphorylation of ERK, prevents microglial activation, greatly reduces apoptotic cells, and does not affect the phosphorylation of tau. Our data suggest that the tau hyperphosphorylation and apoptosis triggered by A beta are two independent events, and that the neuroprotective effect of GDNF against A beta may result either directly by the inhibition of ER stress or indirectly through the inhibition of JNK and ERK activation.

Preen gland function in layer fowls: factors affecting morphology and feather lipid levels.

The effects of age and external factors on preen gland morphology and feather lipid concentrations in layer fowl (Gallus domesticus) have not previously been described. This work examines how floor substrate, whether a bird is feather pecked or not, and age influence both gross and histological morphology. Furthermore, the effects of age, beak trimming and floor substrate on feather lipid concentration (mg lipid per g feathers) are investigated. Neither absolute nor relative preen gland weight were reliably affected by floor substrate or feather pecked status. However, these parameters were strongly affected by changes in bird age and related body weight changes. The microstructures of the gland also increased consistently with age. Feather lipid concentration generally increased with age. Feather lipid was strongly influenced by the presence or absence of a dustbathing substrate. Of all the parameters tested here, the age and growth of the bird had the greatest effects on preen gland development. Feather lipid concentration is also influenced by age, and by the presence or absence of a dustbathing substrate.

MPP+ induces the endoplasmic reticulum stress response in rabbit brain involving activation of the ATF-6 and NF-kappaB signaling pathways.

Inhibition of mitochondrial function and the subsequent generation of oxidative stress are strongly suggested to underlie MPTP/MPP+-induced neurotoxicity, which has been used extensively as a model for Parkinson disease. In the present study we have examined the hypothesis that MPP+ targets the endoplasmic reticulum. Because rabbits possess more genetic similarities to primates than to rodents we have selected this animal model system for our MPP+ neurotoxicity studies. MPP+ was administered directly into the brain of New Zealand white rabbits via the intracisternal route, and the effects on tissue from the substantia nigra were examined. Here we demonstrate that MPP+ in a dose-dependent manner induces the loss of tyrosine hydroxylase activity, oxidative DNA damage, and activation of the endoplasmic reticulum stress response. The endoplasmic reticulum response, mediated by activation of ATF-6 and NF-kappaB, leads to activation of gadd 153. These effects correlate with the activation of caspase-3 and of c-Jun N-terminal kinase (JNK) kinase. We propose that pharmacological agents that inhibit the perturbation of endoplasmic reticulum function or the activation of JNK may represent a potential therapeutic approach for the prevention of neurotoxin-induced Parkinson disease.

Abeta(1-42)-induced JNK and ERK activation in rabbit hippocampus is differentially regulated by lithium but is not involved in the phosphorylation of tau.

Administration of Abeta(1-42) into the rabbit brain induces apoptosis and phosphorylation of tau. These Abeta effects correlate with the activation of JNK and ERK, but not of p38. Treatment with 7 mM lithium inhibits apoptosis, modulates JNK and ERK and does not affect the phosphorylation of tau. Our results demonstrate that lithium, at this dose, effectively inhibits the Abeta-induced apoptosis but has no effect on tau phosphorylation, and that MAP kinases are not involved in the phosphorylation of tau.

Intracellular mechanisms underlying aluminum-induced apoptosis in rabbit brain.

Loss of neurons is a hallmark of neurodegenerative disorders and there is increasing evidence suggesting that apoptosis is a key mechanism by which neurons die in these diseases. Mitochondrial dysfunction has been implicated in this process of neuronal cell death, but there is a growing body of evidence suggesting also an active role for the endoplasmic reticulum in regulating apoptosis, either independent of mitochondria, or in concert with mitochondrial-initiated pathways. Investigations in our laboratory have focused on neuronal injury resulting from the administration of aluminum maltolate, via the intracisternal route, to New Zealand white rabbits. This treatment induces both mitochondrial and endoplasmic reticulum stress. Agents such as lithium or glial cell-line derived neurotrophic factor (GDNF) have the ability to prevent aluminum-induced neuronal death by interfering with the mitochondrial and/or the endoplasmic reticulum-mediated apoptosis cascade. Cytochrome c release from mitochondria and binding to Apaf-1 initiates the aluminum-induced apoptosis cascade; this is prevented by lithium treatment. GDNF also protects against aluminum-induced apoptosis but by upregulation of Bcl-X(L), thereby preventing the binding of cytochrome c to Apaf-1. This animal model system involving neurotoxicity induced by an aluminum compound provides new information on mechanisms of neurodegeneration and neuroprotection.

Lithium inhibits Abeta-induced stress in endoplasmic reticulum of rabbit hippocampus but does not prevent oxidative damage and tau phosphorylation.

The goal of this study was to assess the in vivo effect of Abeta on apoptosis pathways involving the endoplasmic reticulum and mitochondria, and its relationship to the induction of tau phosphorylation and DNA oxidative damage. In rabbits treated intracisternally with aggregated Abeta(1-42), clear evidence of endoplasmic reticulum stress was observed by the activation of caspase-12 and cleavage of caspase-3 in the endoplasmic reticulum. Mitochondrial injury was evident from the release of cytochrome c into the cytosol and the induction of oxidized mitochondrial DNA. Tau phosphorylation and nuclear translocation of NF-kappaB and GSK-3beta were also observed. Treatment with lithium, an inhibitor of GSK-3beta, inhibited caspase activation but did not prevent mitochondrial DNA damage or tau hyperphosphorylation, suggesting that the translocation of GSK-3beta may represent an upstream event that leads to caspase activation but is unrelated to tau hyperphosphorylation or mitochondrial DNA oxidative damage. We propose that treatment by lithium alone is not sufficient to protect against the multiple adverse effects of Abeta, and the use of agents that prevent oxidative DNA damage and tau hyperphosphorylation, together with lithium, may provide better protection from the neurotoxic effect of Abeta.

Massive caffeine overdose requiring vasopressin infusion and hemodialysis.

INTRODUCTION: Massive caffeine overdose is associated with life-threatening hemodynamic complications that present challenges for clinicians. We describe the highest-reported serum concentration of caffeine in a patient who survived and discuss the first-reported use of vasopressin and hemodialysis in a caffeine-poisoned patient. CASE REPORT: A 41-yr-old woman presented 3 h after ingesting approximately 50 g of caffeine. She subsequently underwent cardiopulmonary resuscitation and received multiple medications in an attempt to raise her blood pressure and control her heart rate without success. Vasopressin infusion increased her blood pressure to the point where hemodialysis could be performed. Despite ensuing multisystem organ failure, she survived and has made a complete recovery. CONCLUSION: Hemodialysis and vasopressin infusions may be of benefit in the management of caffeine-intoxicated patients who fail to respond to standard therapies.

The significance of environmental factors in the etiology of Alzheimer's disease.

The proposition that environmental agents, such as diet, aluminum, and viruses, are as important as genetic factors in the etiology of Alzheimer's disease (AD) was advanced by the authors at the Challenging Views of Alzheimer's Disease meeting held in Cincinnati on July 28 and 29, 2001. Diet, dietary fat, and to a lesser extent, total energy (caloric intake), were found to be significant risk factors for the development of AD in a dozen countries, while fish consumption was found to be a significant risk reduction factor. An acid-forming diet, such as one high in dietary fat or total energy, can lead to increased serum and brain concentrations of aluminum and transition metal ions, which are implicated in oxidative stress potentially leading to the neurological damage characteristic of AD. Many of the risk factors for AD, such as cholesterol and fat, and risk reduction factors, such as whole grain cereals and vegetables, are shared with ischemic heart disease. Aluminum may cause neurological damage and a number of studies have linked aluminum to an increased risk for developing AD. The evidence for viral agents playing a role in AD is the strong association between the presence of HSV1 in brain and carriage of an apoE-epsilon4 allele in the case of AD patients but not of controls; statistical analysis shows the association is causal. Diet, aluminum, and viral infections may increase the prevalence of AD by eliciting inflammation, which may cause the neurological damage that results in AD.

Lithium inhibits aluminum-induced apoptosis in rabbit hippocampus, by preventing cytochrome c translocation, Bcl-2 decrease, Bax elevation and caspase-3 activation.

A variety of studies on neuronal death models suggest that lithium has neuroprotective properties. In the present investigation, we have examined the effect of chronic lithium treatment on hippocampus, as monitored by changes at the subcellular level of apoptosis-regulatory proteins which have been induced by the neurotoxin, aluminum maltolate. Intracisternal administration of aluminum into rabbit brain induces cytochrome c release, decreases levels of the anti-apoptotic proteins Bcl-2 and Bcl-X(L), increases levels of the pro-apoptotic Bax, activates caspase-3, and causes DNA fragmentation as measured by the TUNEL assay. Pretreatment for 14 days with 7 mm of lithium carbonate in drinking water prevents aluminum-induced translocation of cytochrome c, and up-regulates Bcl-2 and Bcl-X(L,) down-regulates Bax, abolishes caspase-3 activity and reduces DNA damage. The regulatory effect of lithium on the apoptosis-controlling proteins occurs in both the mitochondria and endoplasmic reticulum. We propose that the neuroprotective effect of lithium involves the modulation of apoptosis-regulatory proteins present in the subcellular organelles of rabbit brain.

The endoplasmic reticulum is the main site for caspase-3 activation following aluminum-induced neurotoxicity in rabbit hippocampus.

We have assessed the distribution of caspase-3 in subcellular fractions from rabbit brain hippocampus and find that in controls the pro-caspase-3 form is distributed mainly in the cytoplasm. In animals treated intracisternally with the neurotoxin aluminum-maltolate, although pro-caspase-3 levels are higher in the cytosolic fractions, p17, the active caspase-3, is localized mainly in the endoplasmic reticulum. This distribution is confirmed by immunohistochemistry which demonstrates the co-localization of p17 with calnexin, a specific marker of the endoplasmic reticulum. Based on the apparent translocation into the endoplasmic reticulum of active caspase-3, an executioner of cell death, the results suggest that this organelle is an important site in the caspase-3 mediated apoptosis cascade. Inhibition of the latter enzyme by directly targeting its main site of activation could represent a strategy to prevent this adverse event.