Molecular Biomarkers & Diagnosis

The etiology of autism spectrum disorders (ASD) is not well known but recently we reported that the serum levels of sonic hedgehog (SHH) protein and brain-derived neurotrophic factor (BDNF) might be linked to oxidative stress in ASD. We hypothesized that Indian hedgehog (IHH) protein which belongs to SHH family may play a pathological role in the ASD. We studied recently diagnosed patients in early stages of ASD (n=54) and age-matched, cognitively normal, individuals (n=25), using serum levels of IHH protein. We found statistically significantly higher-levels of serum IHH protein in ASD subjects (p=0.001) compared to control subjects. Our findings are the first to report a role of IHH in ASD children, suggesting a possible pathological role-played by IHH in early-stage in ASD. Such measures might constitute an early biomarker for ASD and ultimately offer a target for novel biomarker-based therapeutic interventions.

Introduction: The presence of NA against EM regarding specificity and have gained increasing attention in proteome analysis for diagnosis, developing, monitoring and effective treatment of osteoarthritis of the knee (kOA). An understanding of the various regulatory systems controlling blood vessel growth, inflammation and pain in the join should lead to help explain kOA disease progression. To investigate the specific presence of NA (IgM, IgG, IgA) against EM (BK, AII, VEGF, bFGF) in the sera of kOA patients and control and to correlate this with process of joint destruction.

Methods: In this study novel immunoconjugates were designed, synthesized and then used to develop a rapid, specific and sensitive ELISA method to directly detect immune complexes (NA-EM) in humans. Following this procedure, we examined variations in the levels of natural antibodies recognized a panel of self-antigens in the sera from healthy individuals and kOA patients. Blood samples were obtained from 250 patients with symptomatic kOA and 250 ages, sex-matched healthy individuals.

Results: NA against EM was detected with novel ELISA assay in the sera of kOA patients as well as in the sera of control. At time of inclusion kOA patients (100%) had significantly higher BK-IgG levels relative to normal sera. The over expression BK-IgG were positively associated with destructive changes (KL>4; r=0.75; p<0.005). KOA patients in whom KL scores progress rapidly tend to have higher BK-IgG levels at all time point. Serum BK-IgG over expression in kOA patients were positively associated with destructive changes (KL>4; r=0.75; p<0.005). Elevated BK-IgG was significantly correlated with VAS (r=0.85; p<0.0001) and loss of functions (r=0.69; p<0.0003) in kOA patients. Affinity chromatography yielded EM-specific NA from the sera of healthy individuals and kOA patients.

Conclusions: We showed that EM represents a group of novel self-antigens which are targeted by NA from kOA patients. Circulating BK-IgG in the sera has been proposed as a sensitive and specific marker of diagnosing kOA at early stages of the disease. Our results have potential applications for controlling unwanted angiogenesis, inflammation, pain and future response to therapy in kOA patients.

The objective of this review was to focus on recent studies indicating how deregulation of lipid metabolism may be of particular importance for central nervous system (CNS) injuries and neurodegenerative disorders. Furthermore, since an accumulation of neutral lipids (NLs), mainly cholesterol esters (CEs) in the form of cytoplasmic lipid droplets was previously found by our group in peripheral blood mononuclear cells (PBMCs) of Alzheimer (AD) patients and their first degree relatives (AD-FDR), we reviewed current data providing evidence that altered lipid metabolism in brain can also affect cholesterol metabolism in the systemic circulation. Using data from literature we proposed a mechanistic model that helps us to explain why subjects with neurological disorders often accumulate NLs in their PBMCs. If validated by future research, it should provide a rationale for NL-PBMCs determination by Oil Red O (ORO) staining method as a useful tool for diagnostic and therapeutic interventions in AD and possibly in other forms of dementia occurring in childhood as well as in elderly.

Decabromodiphenyl ether (BDE 209) was one of the flame retardants in a variety of commercial and household products. The toxicity of BDE 209 was thought to be associated with neurotoxicity, changes in fetal development, and endocrine disruption etc. Although different toxicology effects of BDE 209 exposure had been reported, the information of BDE 209 on Jurkat cells was still insufficient, and the molecular bio-marks were still unknown. In this research, the gene expression profiling was analyzed. Changes of gene expression induced by BDE 209 could be classified into neurotoxicity, apoptosis and reproductive toxicity. Quantifying microRNA expression levels was verified by real-time PCR. The expression levels of Brain-Derived Neurotrophic Factor (BDNF), Regulating synaptic membrane exocytosis 3 (RIMS3) and dihydropyrimidinase-like 3 (DPYSL3), which were associated with the neurotoxicity and brain development, presented trends of down-regulated. Based on the abundance of gene expression analysis in tissues and cells and taking the possibility of biological samples obtained for the clinical diagnosis into consideration, BDNF, that had been confirmed as one of the key genes in the neurotoxicity, was recommended as one of the main clinical bio-marks in the epidemiology, since it had been found abundant in the blood monocyte.

Adiponectin is a naturally occurring, active protein produced by white adipose tissue circulating levels of which have been associated with breast cancer risk. The ADIPOQ +45T/G and 276G/T polymorphisms are likely to play an important role in the susceptibility to breast cancer. To evaluate the prevalence of polymorphism of Adiponectin gene a case-control study was performed in 154 breast cancer patients and 142 controls in South India. We utilized PCR- RFLP based assay to evaluate the association between the +45T/G and 276G/T polymorphism of the ADIPOQ gene and breast cancer risk in a case control study. Frequencies of ADIPOQ +45T/G were 74%, 24% and 2% in the breast cancer patients and 83%, 15% and 2% in the controls, respectively. ADIPOQ +45T/G genotype showed a 1.7- fold increased risk for breast cancer and the 276G/T genotype showed a 1.6-fold increased risk for breast cancer. In the compound genotypes, T45G//G276T and T45G/T276T showed a 1.668 and a 1.791-fold increased risk for breast cancer. Our findings suggest that the ADIPOQ +45T/G and 276G/T polymorphism may be a useful biomarker associated with the risk of breast cancer in obese women in Indian population

The study evaluated lung damages caused by in vivo exposure to silica nanoparticle doped with cadmium (SiNPs-Cd, 1 mg/rat) in terms of oxidative stress induction, apoptosis, and fibrosis, and assessed the validity of plasma F2-isoprostanes (F2-IsoPs) as marker of pulmonary insult. SiNPs-Cd effect was assessed 24 hr, 7 and 30 days post-intratracheal instillation compared to that caused by CdCl2 (400 μg/rat), or SiNPs (600 μg/rat) characterizing pulmonary superoxide dismutase (SOD1), cyclooxygenase type-2 (COX2) and collagen expression (by immunohistochemistry and TEM), and investigating apoptosis (TUNEL staining). Free and esterified F2- IsoPs were measured in lung and plasma by gas chromatography/negative ion chemical ionization tandem mass spectrometry (GC/NICI-MS/MS) analysis. Lung: SiNPs-Cd induced enhancement of SOD1 and COX2 immunoreactivity in a time-dependent manner (7<30 days). Total F2-IsoPs also increased 30 days post-exposure (46.7 ± 11 ng/g in SiNP-Cd vs. 32.8 ± 7.8 ng/g in control). Parallely, apoptosis enhanced as following SiNPs-Cd>CdCl2>SiNPs. A strong fibrotic response, i.e. interstitial type I collagen over-expression, was also observed starting at 7 days, particularly after SiNPs-Cd. Plasma: Pronounced elevation of free F2-IsoPs occurred (54.6 ± 2 vs. 28 ± 8 pg/ml in SiNPs-Cd andcontrol, respectively) already at day 7 lasting until day 30. In SiNPs-treated animals no changes were observed on oxidative stress parameters. The CdCl2 pulmonary response was milder than that found with SiNPs-Cd. The results indicate long-lasting tissue injury following SiNPs-Cd pulmonary exposure in rat and a role for plasma F2-IsoPs as a predictive indicator of nanoparticle-induced oxidative insult