Molecular Biomarkers & Diagnosis

It is known, human cells are incapable of synthesizing the N-glycolyl neuraminic acid (NeuGc) due to the inactivation of the cytidine monophospho-N-acetyl-neuraminic acid hydroxylase, the enzyme responsible for the synthesis of this sialic acid. Conversely, the aberrant expression of NeuGc-sialoconjugates has been detected in humans, although preferentially in malignant tissues. The more accepted hypothesis for the presence of NeuGc in human malignancies is associated with its incorporation from dietary sources to the altered metabolism of malignant cells which is also favored by the hypoxic conditions of tumors. However, the significance of NeuGccontaining conjugates in tumor biology as well as its clinical implications is still under investigation. One of these molecules is Nglycolyl GM3 ganglioside (NeuGcGM3).

We have designed a standardized protocol with multiplex-primer sets capable of detecting majority IG kappa (IGK) and IG lambda (IGL) light chain rearrangements in plasma cell neoplasms (PCN). Thirty primers were combined in three multiplexed PCR reactions to target IGK, KDE and IGL rearrangements. Variable region (V) primers were designed to prevent “primer dimers”, provide matching melting temperatures (Tm), minimize amplicon size, and optimize sequencing time. Amplicons were subjected to capillary gel electrophoresis for analysis. In a discovery series, we tested 37 plasma cells neoplasms PCN (28 PCNs at diagnosis and 9 PCNs post-treatment). The assay investigated an additional 52 prospective PCN cases in the validation series. Results were compared to bone marrow morphology, immunohistochemical (IHC), flow cytometry data, and standard IGH FRIII gene rearrangement assay. In the discovery series, the following sensitivities/specificities were obtained for mature B-cell neoplasms: IGH FRIII: 29.7%/100%, IGK: 80.4%/100%, KDE: 25.0%/100%, and IGL: 35.1%/96.8%. The combination of IGH FRIII, IGK, and KDE detected 83.8% (31/37) vs 67.3% (35/52) in the discovery vs validation series, respectively, for the PCN population. Interestingly, 21.2% (11/52) of the validation samples positive by IG clonality, were negative by IHC and flow cytometry. In IHC/flow cytometry positive cases with a PCN representing a tumor burden of >50%, 10% to 50%, 1% to 10%, 0% to 1% of cells, the combined sensitivity of the IG clonality assay was 100% (20/20), 72% (23/32), 53% (10/19) and 20% (1/5) respectively. This IGK/IGL clonality assay has good sensitivity at diagnosis.

Particular MYBPC3. A 25-basepair deletion variant of MYBPC3 occurs at high frequency in individuals of South Asian descent and is estimated to affect 55 million people worldwide, carrying an increased likelihood of cardiomyopathy. Since this variant is prevalent and severe in this subpopulation, quick and affordable screening to provide risk-assessment to guide treatment for these patients is critical. An RNaseH qPCR assay was developed to quickly and specifically diagnose the presence of the 25-basepair deletion variant in MYBPC3. RNAseH-blocked nucleotide primers were designed to identify the presence or absence of the wild type MYBPC3 allele or the genomic sequence containing the 25-basepair deletion. Using this assay, three blinded operators were able to accurately determine the genotype from human genomic DNA samples from blood and saliva using a qPCR thermocycler. Furthermore, positive variant subjects were examined by both electrocardiography and echocardiography for the presence of cardiomyopathy. A simple, robust assay was established, verified and validated that can be automated to detect the presence of the highly prevalent 25-basepair deletion MYBPC3 variant using both blood and saliva samples. The assay will provide quick and accurate prescreening of individuals at high risk for cardiomyopathies and allow for better clinical identification of 25-basepair deletion MYBPC3 carriers in large cohort epidemiological studies.

A novel two-photon fluorescence probe for Hg2+ derived from bis(styryl)terephthalonitrile as a two-photon fluorophore and bis [2-(2-hydroxyethyl sulfanyl) ethyl] amino group (ionophore) as a novel Hg2+ ligand was developed. The probe possesses small molecule size, large two-photon absorption cross-section (1067 GM) in H2O, noncytotoxic effect, long-wavelength emission at 588 nm, large Stokes shift (121 nm), excellent photostability, high water-solubility, good cell-permeability, and pH-insensitivity in the biologically relevant range. The probe can selectively detect Hg2+ ions in live cells and living tissues without interference from other metal ions and the membrane-bound probes, and its quenching constant (Ksv TP) is 8.73 × 105 M-1.

Many targeted therapies have been developed to treat lung cancer. Unfortunately, however statistical data over the past two decades suggest only a slight improvement in a patient's survival rate after diagnosis. Clonal evolution and tumor heterogeneity are the major obstacles in designing effective targeted treatments against cancer. To create more comprehensive treatments, emerging therapies target bioenergetic pathways of cancer cells. Like normal cells, cancer cells can generate energy only through glycolysis and oxidative phosphorylation. Notably, a number of studies have shown that many types of cancer cells rely heavily on mitochondrial respiration. Importantly, research carried out in the authors’ laboratory showed that non-small cell lung cancer cells exhibit increased levels of mitochondrial and heme function. Hence, limiting heme availability interferes with bioenergetics of cancer cells. Evidently, targeting heme function may provide an effective way for treating lung cancer.

On the basis of this axioms we are proposing that consciousness is the result of a global workspace in the brain, which distributes information to the huge number of paralell unconscious processors forming a rest of the brain. Our theory is founded on the view that the brain is composed of many different paralell processors, or moduls, each capable of performing some task on the symbolic representations that it receives as input. The moduls are flexible in that they can combine to form new processors capable of performing novel tasks, and can decompose into a smaller component processors. Axiom A: The consciousness is a mathematical structure, with neurobiological semantics. Axiom B: The higher mental processes all correspond to well-defined but, at present, poorly understood information processing functions that are carried out by physical systems, our brains.

Nutrients are partially bioaccessible to be extracted from their matrix. They may be partially lost by processing to a meal and by metabolism after absorption from the GI tract. Numerous steps in this cascade have an impact on the nutrient amount which will finally be bioavailable for an effect. Daily allowances are defined and widely recommended to obtain such an effect. However, these needs are derived from observation of healthy populations and are increased to cover needs for special consumer and patient groups or for pregnancy. Daily allowances are subject to frequent changes whenever new scientific evidence arises as is the case for vitamin D. This article describes reasons why generally reasonable recommendations may fail occasionally are found among the variation of genes, age, gender, or race, as well as epigenetics, which all contribute to the individual expression of metabolic capacity and thus to differences in individual bioavailability.