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Recurrent pregnancy loss (RPL) affects a large number of couples worldwide, increasing their mental and financial burdens. While female factors that contribute to RPL have been studied extensively, the role of male factors is largely unknown, and approximately 40 % RPL cases remain unexplained despite thorough clinical examinations. These cases are clinically termed as idiopathic RPL (iRPL). Several studies have recently found that spermatozoa play an important role, beyond fertilization, in iRPL, specifically in early embryonic development. Consequently, scientists explored spermatozoa to understand iRPL and revealed that both oxidative stress and DNA fragmentation contribute to RPL. In this study, we analyzed sperm samples from male partners of iRPL patients and fertile men who recently fathered a child by LC-MS/MS to identify proteomic markers of iRPL. A total of 1,988 proteins were quantified by a label-free method, and stringent statistical analysis was performed for the selection of candidate biomarkers of iRPL. Out of 1,647 proteins quantified, only 7 proteins qualified the selection criteria, which are lactotransferrin, ATP synthase subunit beta mitochondrial, fatty acid synthase, anterior gradient protein 2 homolog, hemoglobin subunit beta, short-chain specific acyl-CoA dehydrogenase mitochondrial, cytoplasmic dynein 1 heavy chain, and 14-3-3 protein sigma. We then performed gene annotations, pathways, and network analyses to gain more biological insights, identifying an association between oxidative stress and iRPL.

Background Many studies reported for estimating serum YKL-40 using ELISA or RIA methods. This study introduces the plausible utilization of real-time surface plasmon resonance (SPR) technology in investigating the expression of serum YKL-40 protein levels and ELISA method for serum IgE in bronchial asthma. Methods A commercially available BIAcore 2000 instrument, based on SPR technology, was utilized for assessing serum YKL-40 levels in a control sample size of 45 and active sample size of 97. Antibody immobilization was optimized to obtain the best sensor performance and a sensitive analytic detection. A commercially available ELISA kit was utilized for detecting serum IgE to estimate allergic condition-associated asthma. Results The results of SPR technology could distinctly classify with highly statistical significance, the asthma severities by estimating the elevated levels of YKL-40 in blood sera of minute quantities (up to 0.33 ng/ml), and thus differentiates superior utility in comparison with ELISA method. No statistically significant correlation of YKL-40 and IgE was observed. Conclusions Serum YKL-40 may be used as a protein marker in classifying asthma severity by applying SPR technology as a reliable, label-free, highly sensitive, and cost-effective tool.

The currently prescribed tests for asthma diagnosis require compulsory patient compliance, and are usually not sensitive to mild asthma. Development of an objective test using minimally invasive samples for diagnosing and monitoring of the response of asthma may help better management of the disease. Raman spectroscopy (RS) has previously shown potential in several biomedical applications, including pharmacology and forensics. In this study, we have explored the feasibility of detecting asthma and determining treatment response in asthma patients, through RS of serum. Serum samples from 44 asthma subjects of different grades (mild, moderate, treated severe and untreated severe) and from 15 reference subjects were subjected to Raman spectroscopic analysis and YKL-40 measurements. The force expiratory volume in 1 second (FEV1) values were used as gold standard and the serum YKL-40 levels were used as an additional parameter for diagnosing the different grades of asthma. For spectral acquisition, serum was placed on a calcium fluoride (CaF2) window and spectra were recorded using Raman microprobe. Mean and difference spectra comparisons indicated significant differences between asthma and reference spectra. Differences like changes in protein structure, increase in DNA specific bands and increased glycosaminoglycans-like features were more prominent with increase in asthma severity. Multivariate tools using Principal-component-analysis (PCA) and Principal-component based-linear-discriminant analysis (PC-LDA) followed by Leave-one-out-cross-validation (LOOCV), were employed for data analyses. PCA and PC-LDA results indicate separation of all asthma groups from the reference group, with minor overlap (19.4%) between reference and mild groups. No overlap was observed between the treated severe and untreated severe groups, indicating that patient response to treatment could be determined. Overall promising results were obtained, and a large scale validation study on random subjects is warranted before the routine clinical usage of this technique.