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New tumor markers: CA125 and beyond.

A variety of biomarkers have been developed to monitor growth of ovarian cancer and to detect disease at an early interval. CA125 (MUC16) has provided a useful serum tumor marker for monitoring response to chemotherapy, detecting disease recurrence, distinguishing malignant from benign pelvic masses, and potentially improving clinical trial design. A rapid fall in CA125 during chemotherapy predicts a favorable prognosis and could be used to redistribute patients on multiarmed randomized clinical trials. Several studies now document that CA125 can serve as a surrogate marker for response in phase II trials. Serial measurement of CA125 might also provide a useful marker for monitoring stabilization of disease with cytostatic targeted therapeutic agents. The greatest potential for serum markers may be in detecting ovarian cancer at an early stage. A rising CA125 can be used to trigger transvaginal sonography (TVS) in a small fraction of patients. An algorithm has been developed that calculates risk of ovarian cancer based on serial CA125 values and refers patients at highest risk for TVS. Use of the algorithm is currently being evaluated in a trial with 200,000 women in the UK that will test critically the ability of a two-stage screening strategy to improve survival in ovarian cancer. Whatever the outcome, as 20% of ovarian cancers have little or no expression of CA125, additional serum markers will be required to detect all patients in an initial phase of screening. More than 30 serum markers have been evaluated alone and in combination with CA125 by different investigators. Some of the most promising include: HE4, mesothelin, M-CSF, osteopontin, kallikrein(s), and soluble EGF receptor. Two proteomic approaches have been used: one examines the pattern of peaks on mass spectroscopy and the other uses proteomic analysis to identify a limited number of critical markers that can be assayed by more conventional methods. Both approaches are promising and require further development. Several groups are placing markers on multiplex platforms to permit simultaneous assay of multiple markers with very small volumes of serum. Mathematical techniques are being developed to analyze combinations of marker levels to improve sensitivity and specificity. In the future, serum markers should improve the sensitivity of detecting recurrent disease as well as facilitate earlier detection of ovarian cancer.

Biomarkers, Tumor↗

Ozone/biotic disease interactions: molecular biomarkers as a new experimental tool.

Current climate change scenarios predict a further increase of tropospheric ozone which is well known to inhibit plant photosynthesis and growth processes. Ozone can also predispose plants to enhanced biotic attack, as proposed in particular for necrotrophic fungi, root-rot fungi and bark beetles. However, at present it does not seem possible to predict whether increased ambient ozone will lead to a higher or lower disease likelihood in particular plant-pathogen systems. It has been stated repeatedly in the literature that periods of high ambient ozone are essentially non-coincident with infection periods of most fungal pathogens. This implies minimal interactive risks. However, it now appears that the various ozone-induced metabolic changes can persist in plants over days or months. Visible ozone symptoms also may be greatly delayed. Certain stress transcripts, proteins and metabolites have been developed as ozone biomarkers in controlled exposure experiments, but these biomarkers remain to be examined on field sites. A simple epidemiological scenario based on 'memory' time spans of ozone effects is proposed as a tool to make ozone-plant disease interactions more predictable.

Journal Article↗

[Serum gamma-glutamyltransferase levels and the risks of impaired fasting glucose in healthy men: a 2-year follow-up].

OBJECTIVES: An increase in the serum gamma-glutamyltransferase (GGT) concentration has been regarded as a marker of alcohol drinking or liver disease. Some reports, however, have suggested that the serum GGT may be a sensitive and early biomarker for the development of prediabetes and diabetes. In this study we investigated whether serum GGT is a reliable predictor of the incident impaired fasting glucose (IFG), including diabetes. METHODS: We performed a prospective study for two years (2002-2004). We analyzed the periodic health examination data from a total of 4,711 men. The examinations were done in the years 2002 and 2004. The analyzed data included a self-questionnaire, a physical examination and the laboratory results. Both IFG and diabetes were defined as a serum fasting glucose concentration of more than 100 mg/dL and 126 mg/dL, respectively. RESULTS: A total of 738 cases (15.7%) of incident IFG and 13 cases (0.3%) of diabetes occurred. The mean serum GGT concentrations were quite different between the normal (38.0 IU) and incident IFG groups (50.3 IU), and the incident diabetes group (66.0 IU) (p <0.001). After multivariable adjustment, the relative risks for incident IFG or diabetes across the baseline GGT categories (<10th, 10th-20th, 30th-40th, 50th-60th, 70th-80th and >90th percentile) were 1.0, 1.172 (0.769-1.785), 1.107 (0.725-1.689), 1.444 (0.934-2.232), 2.061 (1.401-3.031) and 2.545 (1.784-3.631) (p-value for trend: <0.001). The risks significantly increased with increasing levels of GGT for 2 years; when comparing the increased groups (<10%, .10-20%, >20%) versus the decreased over 20% group of GGT, the risks for IFG or diabetes were 1.334 (1.002-1.776), 1.613 (1.183-2.199) and 1.399 (1.092-1.794). CONCLUSIONS: Our findings suggest that serum GGT concentrations within its normal range may be an early predictor of the development of IFG and diabetes. As serum GGT is a relatively inexpensive test and a reliable marker, it might have important implications in public health promotion.

Biomarkers↗

Microarray analysis identifies keratin loci as sensitive biomarkers for thyroid hormone disruption in the salamander Ambystoma mexicanum.

Ambystomatid salamanders offer several advantages for endocrine disruption research, including genomic and bioinformatics resources, an accessible laboratory model (Ambystoma mexicanum), and natural lineages that are broadly distributed among North American habitats. We used microarray analysis to measure the relative abundance of transcripts isolated from A. mexicanum epidermis (skin) after exogenous application of thyroid hormone (TH). Only one gene had a >2-fold change in transcript abundance after 2 days of TH treatment. However, hundreds of genes showed significantly different transcript levels at days 12 and 28 in comparison to day 0. A list of 123 TH-responsive genes was identified using statistical, BLAST, and fold level criteria. Cluster analysis identified two groups of genes with similar transcription patterns: up-regulated versus down-regulated. Most notably, several keratins exhibited dramatic (1000 fold) increases or decreases in transcript abundance. Keratin gene expression changes coincided with morphological remodeling of epithelial tissues. This suggests that keratin loci can be developed as sensitive biomarkers to assay temporal disruptions of larval-to-adult gene expression programs. Our study has identified the first collection of loci that are regulated during TH-induced metamorphosis in a salamander, thus setting the stage for future investigations of TH disruption in the Mexican axolotl and other salamanders of the genus Ambystoma.

Ambystoma↗

Estimation of the day-specific probabilities of conception: current state of the knowledge and the relevance for epidemiological research.

Conception, as defined by the fertilisation of an ovum by a sperm, marks the beginning of human development. Currently, a biomarker of conception is not available; as conception occurs shortly after ovulation, the latter can be used as a proxy for the time of conception. In the absence of serial ultrasound examinations, ovulation cannot be readily visualised leaving researchers to rely on proxy measures of ovulation that are subject to error. The most commonly used proxy measures include: charting basal body temperature, monitoring cervical mucus, and measuring urinary metabolites of oestradiol and luteinising hormone. Establishing the timing of the ovulation and the fertile window has practical utility in that it will assist couples in appropriately timing intercourse to achieve or avoid pregnancy. Identifying the likely day of conception is clinically relevant because it has the potential to facilitate more accurate pregnancy dating, thereby reducing the iatrogenic risks associated with uncertain gestation. Using data from prospective studies of couples attempting to conceive, several researchers have developed models for estimating the day-specific probabilities of conception. Elucidating these will allow researchers to more accurately estimate the day of conception, thus spawning research initiatives that will expand our current limited knowledge about the effect of exposures at critical periconceptional windows. While basal body temperature charting and cervical mucus monitoring have been used with success in field-based studies for many years, recent advances in science and technology have made it possible for women to get instant feedback regarding their daily fertility status by monitoring urinary metabolites of reproductive hormones in the privacy of their own homes. Not only are innovations such as luteinising hormone test kits and digital fertility monitors likely to increase study compliance and participation rates, they provide valuable prospective data that can be used in epidemiological research. Although we have made great strides in estimating the timing and length of the fertile window, more work is needed to elucidate the day-specific probabilities of conception using proxy measures of ovulation that are inherently subject to error. Modelling approaches that incorporate the use of multiple markers of ovulation offer great promise to fill these important data gaps.

Biomarkers↗

Identification of novel peptide safety markers for exocrine pancreatic toxicity induced by cyanohydroxybutene.

Historically, serum amylase and lipase levels have been used to indicate pancreas injury; however, these enzyme levels are often not predictive of pathology. In an effort to discover novel biomarkers of pancreatic acinar cell injury, we analyzed serum and pancreas tissue from cyanohydroxybutene (CHB)-treated male IGS rats using proteomics methods. CHB produces an "edematous pancreatitis," characterized by depletion of zymogen granules, acinar cell apoptosis, and mild to moderate inflammation. Secondary necrosis occurs at higher doses. Rats were treated with 150 mg/kg of CHB and samples were collected at 4, 8, and 24 h. Analyses of serum tryptic digests by surface-enhanced laser desorption-ionization mass spectrometry revealed two novel peptide biomarkers (RA1609 and RT2864) that were predictive of pancreatic damage. Levels of RA1609 decreased, while levels of RT2864 increased by 8 h following CHB treatment. The changes in RA1609 and RT2864 were detected in media from CHB-treated primary rat acini, demonstrating that these peptides are either of pancreatic cell origin or are produced by proteases released from acinar cells. Sequencing revealed that RA1609 is a fragment of rat albumin (accession number P02770, residues 348-360) and RT2864 is a portion of either rat trypsin III (accession number P08426, residues 39-65) or bovine trypsin (accession number P00760, residues 35-61). These two peptides, and possibly other fragments of serum proteins that are digested by pancreatic proteases, may be useful as safety markers for exocrine pancreatic toxicity during drug development or as biomarkers for the diagnosis and/or grading of severity of pancreatic disease.

Acute Disease↗

Oncogenic PI3K and its role in cancer.

PURPOSE OF REVIEW: The purpose of this review is to examine the contribution of the PI3K signaling pathway to the development of human tumors and to propose further studies to elucidate how to develop therapeutics for patients with mutations in this pathway. RECENT FINDINGS: More than 30% of various solid tumor types were recently found to contain mutations in PIK3CA, the catalytic subunit of PI3K. Further analysis of key genes in this pathway identified an additional eight genes altered in tumors. These were generally found to be mutated in a mutually exclusive manner, thus increasing the mutation frequency of the pathway to 40% in colorectal cancers and emphasizing the importance of the PI3K pathway in tumorigenesis. Functional analyses of PIK3CA mutations revealed that they increase its enzymatic activity, stimulate AKT signaling, allow growth factor-independent growth as well as increasing cell invasion and metastasis. SUMMARY: The PI3K signaling pathway is dysregulated by a variety of mechanisms in a large fraction of human tumors. Both mutational and functional analyses have shown that PIK3CA is an oncogene that plays an important role in tumor progression. Mutant members of the PI3K pathway, including PIK3CA, are good targets for therapeutic intervention because most of them are kinases, making them attractive for drug development. Gaining further insights into PIK3CA oncogenic mechanisms may produce new biomarkers and help the development of targeted therapeutics.

Cell Transformation, Neoplastic↗

Molecular epidemiology in environmental health: the potential of tumor suppressor gene p53 as a biomarker.

One of the challenges in environmental health is to attribute a certain health effect to a specific environmental exposure and to establish a cause-effect relationship. Molecular epidemiology offers a new approach to addressing these challenges. Mutations in the tumor suppressor gene p53 can shed light on past environmental exposure, and carcinogenic agents and doses can be distinguished on the basis of mutational spectra and frequency. Mutations in p53 have successfully been used to establish links between dietary aflatoxin exposure and liver cancer, exposure to ultraviolet light and skin cancer, smoking and cancers of the lung and bladder, and vinyl chloride exposure and liver cancer. In lung cancer, carcinogens from tobacco smoke have been shown to form adducts with DNA. The location of these adducts correlates with those positions in the p53 gene that are mutated in lung cancer, confirming a direct etiologic link between exposure and disease. Recent investigations have also explored the use of p53 as a susceptibility marker for cancer. Furthermore, studies in genetic toxicology have taken advantage of animals transgenic for p53 to screen for carcinogens in vivo. In this review, we summarize recent developments in p53 biomarker research and illustrate applications to environmental health.

Aflatoxin B1↗

Long-term test-retest reliability of functional MRI in a classification learning task.

Functional MRI is widely used for imaging the neural correlates of psychological processes and how these brain processes change with learning, development and neuropsychiatric disorder. In order to interpret changes in imaging signals over time, for example, in patient studies, the long-term reliability of fMRI must first be established. Here, eight healthy adult subjects were scanned on two sessions, 1 year apart, while performing a classification learning task known to activate frontostriatal circuitry. We show that behavioral performance and frontostriatal activation were highly concordant at a group level at both time-points. Furthermore, intra-class correlation coefficients (ICCs), which index the degree of correlation between subjects at different time-points, were high for behavior and for functional activation. ICC was significantly higher within the network recruited by learning than outside that network. We conclude that fMRI can have high long-term test-retest reliability, making it suitable as a biomarker for brain development and neurodegeneration.

Adult↗

Detection of tumor markers with ProteinChip technology.

The early diagnosis of cancer at a curable stage is crucial for the successful treatment of this disease. Most of the currently used tumor assays appear too late and rely on single biomarkers with high false-negative and/or false-positive rates. As an additional burden for the patient, the traditional assays often require biopsy material instead of less invasively taken samples like serum. With the hope for more reliable DNA- and RNA-based screening tools, the research activities of the past 20 years have focused on the genomic characteristics of cancer cells. But, up to now, the output from this strategy has been disappointingly low and the disillusionment is paired with a return to proteins as the real key players in all physiological and pathological processes. Meanwhile, comparative protein profiling is generally acknowledged as a promising way for the detection of specific and predictive protein patterns reflecting certain stages of cancer without dependency on single markers. To meet the new technological demands, the ProteinChip Biomarker System was developed for the Expression Difference Mapping analysis of several hundreds of samples per day on a single, uncomplicated platform; with software support for the construction of multi-marker predictive models. The Interaction Discovery Mapping platform is introduced as the next methodical step for investigations about protein binding partners of possible importance in diagnosis and therapy. This review summarizes the current state in cancer diagnosis, provides an introduction into the ProteinChip technology, and gives an update on publications and research collaborations in SELDI-based tumor marker discovery.

Animals↗

Application of proteomics and protein analysis for biomarker and target finding for immunotherapy.

Regulatory T-cells play a central role in the maintenance of the immunological balance and are powerful inhibitors of T-cell activation both in vivo and in vitro. The enhancement of suppressor-cell function might be a target for immunotherapeutic approaches for the treatment of immune-mediated diseases like multiple sclerosis and Crohn's disease.The method of choice to elucidate the still unclear effector functions of regulatory T-cells is the differential proteome analyses performed with human and murine T-cell populations. To this end, whole-protein extracts of conventional and regulatory T-cells are separated by high-resolution two-dimensional gel electrophoresis according to Klose. The proteomes are analyzed by a 2DE gel image analysis software, ProteomWeaver. The protein spots that are found differentially expressed are picked from the gels and prepared for matrix-assisted laser desorption/ionization (MALDI) mass spectrometrical analysis automatically. The high-resolution 2DE-PAGE and the automated spot handling and protein identification allows one to rapidly find new potential candidate proteins that are of functional relevance for regulatory T-cells, to be used as targets for drug development or as biomarkers for research and diagnostic purposes.

Amino Acid Sequence↗

Effect of potassium dichromate and fenitrothion on hemoglobins of Chironomus riparius Mg. (Diptera, Chironomidae) larvae: potential biomarker of environmental monitoring.

In an attempt to identify Chironomus hemoglobins as biomarkers for environmental monitoring, alterations in the hemoglobins in Chironomus riparius Mg. (Diptera: Chironomidae) larvae, exposed to potassium dichromate and fenitrothion, were investigated under laboratory conditions. The hemoglobins were evaluated in terms of their total contents by a cyanomethemoglobin procedure, individual components by electrophoresis of isoelectric focusing, and their oxidation by multi wavelength rapid-scanning spectrophotometry. The total hemoglobin contents increased at the high level of fenitrothion exposure. No variations in the individual hemoglobin component levels were found, by exposure to either fenitrothion or potassium dichromate. Whereas, the absorption spectra of the hemoglobins showed decreases in the peaks corresponding to the oxyhemoglobins by exposure to both compounds, but more sensitively by the chromium, which probably reflects the increase of the autoxidation of the oxyhemoglobins to methemoglobins by these compounds. These results suggest that autoxidation of the hemoglobins in Chironomus riparius seems to be a sensitive parameter in response to redox-active chemical exposure, and this biochemical parameter could be developed as a biomarker in environmental monitoring.

Animals↗

New technologies and directed agents for applications of cancer imaging.

Molecular imaging represents tissue-specific imaging and quantification of physiologic (functional) and molecular events in tumors utilizing new noninvasive imaging modalities, radioligands, and contrast agents. It combines anatomic, physiologic, and metabolic information in a single imaging session. Molecular imaging relies on the ability to target genes and proteins that are linked directly or indirectly to human disease. New imaging biomarkers are being developed. In addition, functional and molecular imaging can potentially replace anatomic longitudinal studies by assessing treatment response earlier. Vascular targeting agents can be evaluated by imaging of tumor angiogenesis using magnetic resonance imaging (MRI), computed tomography and ultrasound, and positron emission tomography (PET). Targeted contrast agents can accomplish site-directed imaging or therapy by a variety of active and passive mechanisms. Furthermore, there is the possibility of combining different modalities such as ultrasonic imaging and MRI or MRI and PET to increase the flexibility unachievable with either modality alone. However, there is a need to standardize these techniques so that longitudinal evaluation of tumor response to treatment is feasible.

Biomarkers, Tumor↗

DNA as a dietary biomarker in Antarctic krill, Euphausia superba.

The diet of Antarctic krill (Euphausia superba) has been studied using a variety of techniques, but current methods still suffer from problems that are difficult to solve. This study examined an alternative approach utilizing DNA as a prey biomarker. Methods were developed for the preservation, extraction, and identification of prey DNA from krill collected in the field. Group-specific polymerase chain reaction (PCR) was used to amplify diatom prey (Phylum: Bacillariophyta) and the results from DNA clone libraries were compared with microscopic diet analysis. DNA analysis was superior to microscopy for prey detection. However, differences in prey relative abundance estimates between the two techniques suggested some bias in the DNA-based estimates. Quantification showed that large amounts of prey DNA had been successfully preserved and extracted. Overall the results suggest that the application of DNA-based diet analysis to krill warrants further investigation, particularly for prey that are difficult to study using other methods.

Animals↗

Circulating IL-8 and anti-IL-8 autoantibody in patients with ovarian cancer.

OBJECTIVES: In an ongoing effort to identify diagnostic ovarian cancer biomarkers, SEREX (serological analysis of recombinant cDNA expression libraries) technique was employed resulting in detection of 20 known genes, nine ESTs and one novel sequence. Interleukin-8 (IL-8) was one of ovarian cancer-associated antigens identified by SEREX screening. The objective of this study was, therefore, to evaluate the potential importance of circulating anti-IL-8 antibody as ovarian cancer biomarker. METHODS: We developed and optimized a new immunofluorescent bead-based assay for detection of anti-IL-8 antibody in blood serum. Circulating IL-8 and anti-IL-8 IgG concentrations were measured in blood sera from 44 patients with early stage (I-II) ovarian cancer, 50 patients with late stage (III-IV) ovarian cancer, 37 patients with benign pelvic masses, and 80 healthy women using the bead-based assay. RESULTS: Our data indicate that serum contains IL-8 cytokine, anti-IL-8 antibody, and IL-8:anti-IL-8 complexes. We found that concentrations of IL-8 and anti-IL-8 antibody were elevated in sera of patients with ovarian cancer as compared with healthy controls. Logistic regression analysis of circulating concentrations of anti-IL-8 IgG in patients with stages I-II ovarian cancer versus healthy controls allowed for prediction of early ovarian cancer with 98% specificity, 65.5% sensitivity, 80.3% of patients correctly classified. Combining IL-8 and anti-IL-8 IgG with CA 125 resulted in increased classification power as compared to individual markers analyzed separately. CONCLUSION: Thus, IL-8 and anti-IL-8 autoantibody might potentially serve as additional biomarkers for ovarian cancer.

Autoantibodies↗

Lack of an association between the angiotensin-converting enzyme insertion/deletion polymorphism and intracranial aneurysms in a Caucasian population in the United States.

OBJECT: The identification of polymorphisms associated with an increase in the risk of developing disease is integral to the development of genetic biomarkers to identify individuals at risk. Based on reports indicating a role for angiotensin-converting enzyme (ACE) in the pathogenesis of intracranial aneurysms (IAs) as well as hypertension, an independent risk factor for IAs, the authors investigated the association between an insertion/deletion (I/D) polymorphism in the ACE gene and IAs in a Caucasian population in the US. METHODS: The patient population consisted of 162 randomly selected Caucasian patients who underwent surgical repair of an IA at Memorial-Hermann Hospital (Houston, TX) and had no family history of the disease. The ACE I/D polymorphism was typed using polymerase chain reaction amplification of genomic DNA, and allele and genotype frequencies were compared between the patients with IAs and 143 healthy Caucasian volunteers (control group) by performing logistic regression and chi-square tests. The ACE I/D allele frequencies did not differ significantly between the patient and control populations. There were similar allele and genotype frequencies in male and female study participants in both patient and control populations. The authors found no evidence of an association between the allelic or genotypic distribution of the ACE I/D polymorphism and aneurysmal subarachnoid hemorrhage or unruptured IAs. CONCLUSIONS: Contrary to findings in two European Caucasian populations (one British and one Polish), this polymorphism did not contribute to the risk of developing IAs in a Caucasian population in the US.

Adult↗

Proteomic patterns of preinvasive bronchial lesions.

PURPOSE: A proteomics approach is warranted to further elucidate the molecular steps involved in lung tumor development. We asked whether we could classify preinvasive lesions of airway epithelium according to their proteomic profile. EXPERIMENTAL DESIGN: We obtained matrix-assisted laser desorption/ionization time-of-flight mass spectrometry profiles from 10-microm sections of fresh-frozen tissue samples: 25 normal lung, 29 normal bronchial epithelium, and 20 preinvasive and 36 invasive lung tumor tissue samples from 53 patients. Proteomic profiles were calibrated, binned, and normalized before analysis. We performed class comparison, class prediction, and supervised hierarchic cluster analysis. We tested a set of discriminatory features obtained in a previously published dataset to classify this independent set of normal, preinvasive, and invasive lung tissues. RESULTS: We found a specific proteomic profile that allows an overall predictive accuracy of over 90% of normal, preinvasive, and invasive lung tissues. The proteomic profiles of these tissues were distinct from each other within a disease continuum. We trained our prediction model in a previously published dataset and tested it in a new blinded test set to reach an overall 74% accuracy in classifying tumors from normal tissues. CONCLUSIONS: We found specific patterns of protein expression of the airway epithelium that accurately classify bronchial and alveolar tissue with normal histology from preinvasive bronchial lesions and from invasive lung cancer. Although further study is needed to validate this approach and to identify biomarkers of tumor development, this is a first step toward a new proteomic characterization of the human model of lung cancer tumorigenesis.

Aged↗

Detection of biomarkers with a multiplex quantitative proteomic platform in cerebrospinal fluid of patients with neurodegenerative disorders.

Biomarkers are needed to assist in the diagnosis and medical management of various neurodegenerative disorders, including Alzheimer's disease (AD), Parkinson's disease (PD), and dementia with Lewy body (DLB). We have employed a multiplex quantitative proteomics method, iTRAQ (isobaric Tagging for Relative and Absolute protein Quantification), in conjunction with multidimensional chromatography, followed by tandem mass spectrometry (MS/MS), to simultaneously measure relative changes in the proteome of cerebrospinal fluid (CSF) obtained from patients with AD, PD, and DLB compared to healthy controls. The diagnosis of AD and DLB was confirmed by autopsy, whereas the diagnosis of PD was based on clinical criteria. The proteomic findings showed quantitative changes in AD, PD, and DLB as compared to controls; among more than 1,500 identified CSF proteins, 136, 72, and 101 of the proteins displayed quantitative changes unique to AD, PD, and DLB, respectively. Eight unique proteins were confirmed by Western blot analysis, and the sensitivity at 95% specificity was calculated for each marker alone and in combination. Several panels of unique makers were capable of distinguishing AD, PD and DLB patients from each other as well as from controls with high sensitivity at 95% specificity. Although these preliminary findings must be validated in a larger and different population of patients, they suggest that a roster of proteins may be generated and developed into specific biomarkers that could eventually assist in clinical diagnosis and monitoring disease progression of AD, PD and DLB.

Alzheimer Disease↗