Deciphering the potential of proteomic-based biomarkers in women's reproductive diseases: empowering precision medicine in gynecology.

CONTEXT: Gynecological disorders represent a complex set of malignancies that result from a diverse array of molecular changes affecting the lives of over a million women worldwide. Ovarian, Endometrial, and Cervical cancers, Endometriosis, PCOS are the most prevalent ones that pose a grave threat to women's health. Proteomics has emerged as an invaluable tool for developing novel biomarkers, screening methods, and targeted therapeutic agents for gynecological disorders. Some of these biomarkers have been approved by the FDA, but regrettably, they have a constrained diagnostic accuracy in early-stage diagnosis as all of these biomarkers lack sensitivity and specificity. Lately, high-throughput proteomics technologies have made significant strides, allowing for identification of potential biomarkers with improved sensitivity and specificity. However, limited successes have been shown with translation of these discoveries into clinical practice. OBJECTIVE: This review aims to provide a comprehensive overview of the current and potential protein biomarkers for gynecological cancers, endometriosis and PCOS, discusses recent advances and challenges, and highlights future directions for the field. CONCLUSION: We propose that proteomics holds great promise as a powerful tool to revolutionize the fight against female reproductive diseases and can ultimately improve personalized patient outcomes in women's biomedicine.

The Latest Developments in Using Proteomic Biomarkers from Urine and Serum for Non-Invasive Disease Diagnosis and Prognosis.

Due to diagnostic improvements, medical diagnostics is demanding non-invasive or minimally invasive methods. Non-invasively obtained body fluids (eg., Urine, serum) can replace cerebral fluid, amniotic fluid, synovial fluid, bronchoalveolar lavage fluid, and others for diagnostic reasons. Many illnesses are induced by perturbations of cellular signaling pathways and associated pathway networks as a result of genetic abnormalities. These disturbances are represented by a shift in the protein composition of the fluids surrounding the tissues and organs that is, tissue interstitial fluid (TIF). These variant proteins may serve as diagnostic "signatures" for a variety of disorders. This review provides a concise summary of urine and serum biomarkers that may be used for the diagnosis and prognosis of a variety of disorders, including cancer, brain diseases, kidney diseases, and other system diseases. The studies reviewed in this article suggest that serum and urine biomarkers of various illnesses may be therapeutically useful for future diagnostics. Correct illness management is crucial for disease prognosis, hence non-invasive serum and urine biomarkers have been extensively studied for diagnosis, subclassification, monitoring disease activity, and predicting treatment results and consequences.

Colonization of mouse vagina with Pseudomonas aeruginosa: A plausible explanation for infertility.

Little attention has been paid to the influence of asymptomatic colonizers of genital tract on female infertility. Albeit, a variety of uropathogens have been known to negatively alter sperm parameters in vitro, but their impact on female fertility outcome under in vivo conditions is not clearly established. Therefore, the present study was intended to investigate the effect of Pseudomonas aeruginosa on sperm parameters and to identify its role in female infertility. The strain of P. aeruginosa was found to reduce sperm motility, viability and sperm Mg++ATPase activity. It could also lead to premature acrosomal loss and induce morphological defect in spermatozoa. For fertility studies, female mice administered intravaginally with 104, 106, 108 cfu of P. aeruginosa for 10 consecutive days, were allowed to mate with proven breeder male on day 12. The results showed that group of mice receiving P. aeruginosa were rendered infertile whereas group receiving PBS showed abdominal distension, string of pearls and finally delivered pups at the end of gestation period. Further, no other clinical manifestation could be observed apparently, histologically or immunologically. Thus, it can be concluded that infertility in mice might be attributed to asymptomatic colonization of genital tract with sperm immobilizing P. aeruginosa.

Amelioration of sperm immobilisation factor-induced infertility by bacterial antigenic determinants cross-reacting with spermatozoa.

A strain of Staphylococcus aureus, capable of invitro immobilisation of human and mouse spermatozoa, was already present in our laboratory. Therefore, in the present study, the factor responsible (sperm immobilisation factor, SIF) was isolated and purified. It was found to compromise not only motility, but also viability, morphology and Mg2+-ATPase activity of mouse spermatozoa. Also, SIF (250µgmL-1), when administered intravaginally in female BALB/c mice before mating, showed 100% contraceptive effect. Moreover, fluorescein isothiocyanate-labelled SIF was also found to bind mouse spermatozoa and various motile as well as non-motile bacteria, indicating the presence of common SIF-binding receptors on spermatozoa and bacteria. Further, to demonstrate molecular mimicry, the amelioration of SIF-induced impairment of sperm function by a SIF-binding bacterial receptor was compelling. For this, the SIF-binding receptor from Escherichia coli (E-SBR) was purified and evaluated for its ameliorative effect on SIF-induced sperm impairment invitro and invivo. Interestingly, upon the addition of mouse spermatozoa to SIF pre-incubated with E-SBR, an ameliorative effect against SIF-induced impairment of sperm function could be observed through analysis of normal sperm parameters (motility, viability, morphology, Mg2+-dependent ATPase levels). E-SBR also blocked binding of labelled SIF to spermatozoa and bacteria and alleviated SIF-induced infertility in female BALB/c mice. This provided evidence for molecular similarities between bacteria and spermatozoa, owing to which anti-bacterial antibodies cross-reacting with spermatozoa might be produced and infertility might follow.

Receptor from Streptococcus pyogenes as a potential antidote against sperm immobilization factor-induced sperm impairment and infertility.

The present study was undertaken to demonstrate the existence of mimicry between spermatozoa and bacteria. For this, the shared antigenic determinants between mouse spermatozoa and Streptococcus pyogenes against a common ligand, sperm immobilization factor (SIF), were isolated. The mimicry was established on the basis of their ability to ameliorate the SIF-mediated compromised sperm parameters in vitro viz. motility, viability, morphology and Mg2+-ATPase activity of spermatozoa. Further, both the receptors i.e. SIF-binding receptor from mouse spermatozoa (MS-SBR) and SIF-binding receptor from S. pyogenes (S-SBR) were able to block the binding of FITC-labelled SIF to spermatozoa and bacteria. The in vivo studies also showed that MS-SBR (10 µg)/S-SBR (25 µg) could alleviate SIF-induced infertility in female BALB/c mice, further providing evidence for molecular similarities between bacteria and spermatozoa.

Synergism in dual functionality of cryptdin-2 in conjunction with antibiotics against Salmonella.

BACKGROUND & OBJECTIVES: The emergence of multidrug-resistant Salmonella over the last two decades poses a major health risk. In this context, antimicrobial peptides have found a strategic place in the therapeutic armamentarium. Previously, we found that cryptdin-2 has the potential to augment the activity of conventional second- and third-generation anti-Salmonella antibiotics as evident by in vitro assays. In continuation to this, the present study was designed to evaluate the in vivo synergistic effects, if any, of cryptdin-2 in combination with ciprofloxacin and ceftriaxone against murine salmonellosis. METHODS: Scanning electron microscopy (SEM) studies along with in vivo synergistic studies were performed using cryptdin- 2 and antibiotic combinations. In addition, peroxidative liver damage, levels of nitric oxide (NO) and antioxidant enzymes along with tumour necrosis factor-alpha (TNF-α) levels were also measured. RESULTS: The SEM results revealed marked changes on the outer membrane of the bacterial cells treated with various combinations. Both the tested combinations demonstrated synergistic in vivo potency against S. Typhimurium as evident by reduction in the number of Salmonellae in the liver, spleen and intestine. Analysis of peroxidative liver damage, levels of NO and antioxidant enzymes along with TNF-α and nuclear factor-kappa B levels revealed that the tested combinations restored their levels to near normal. The most potent combination was found to be that of cryptdin-2 and ciprofloxacin in terms of direct killing and immunomodulatory potential. INTERPRETATION & CONCLUSIONS: These findings suggest that cryptdin-2 may act in conjunction with conventional antibiotics indicating the possibility of developing these combinations as additional therapeutic agents to combat Salmonella infections.

Impairment by sperm agglutinating factor isolated from Escherichia coli: receptor specific interactions.

In an earlier work done in our laboratory, we have been able to isolate a sperm agglutinating strain of Escherichia coli from the semen sample of a male attending infertility clinic. Further, factor responsible for sperm agglutination (SAF) was isolated and purified, and, using SAF as a tool, corresponding SAF binding receptor from human spermatozoa has been purified. Characterization of SAF and SAF binding receptor using MALDI-TOF showed homology to glutamate decarboxylase and MHC class I molecule, respectively. Coincubation of SAF with spermatozoa not only resulted in spermagglutination but could also compromise other sperm parameters, namely, Mg(2+) dependent ATPase activity and apoptosis. Intravaginal administration of SAF could lead to infertility in Balb/c mice. SAF induced impairment of sperm parameters, and infertility was observed to be due to interaction of SAF with sperm surface receptor component as, when purified receptor was introduced, receptor completely inhibited all the detrimental effects induced by SAF. From these results, it could be concluded that interaction of SAF with spermatozoa is receptor mediated.

Infertility as a consequence of spermagglutinating Staphylococcus aureus colonization in genital tract of female mice.

Various studies have shown Staphylococcus aureus to be one of the most prevalent organism in male and female genital tract but most practitioners dismiss it as mere contamination which is assumed to be of no significance. However, it is now suggested that the presence of this organism should not be ignored, as incubation of spermatozoa with S. aureus results in reduced sperm motility. Although S. aureus has been reported to cause immobilization of spermatozoa, however, its role in infertility has yet to be elucidated. The present study was designed to establish a spermagglutinating strain of S. aureus isolated from the cervix of a woman with unexplained infertility, in mouse and evaluate its effect on fertility outcome. Female Balb/c mice were inoculated intravaginally with different doses of S. aureus (10(4), 10(6) or 10(8)cfu/20 µl) for 10 consecutive days. Microbial colonization monitored every 3(rd) day by vaginal cultures, revealed that strain could efficiently colonize mouse vagina. Mating on day 12, with proven breeder males led to 100% decrease in fertility as compared to control. Even a single dose of 10(6) or 10(8)cfu could lead to vaginal colonization which persisted for 10 days followed by gradual clearing till 21 days, vaginal cultures were negative thereafter. Female mice mated on day 7 (culture positive), were rendered infertile, however, the mice mated on day 22 (culture negative), retained fertility and delivered pups indicating its role in provoking infertility. Further, except infertility, no other clinical manifestation could be seen apparently or histologically. However, when a non-spermagglutinating/immobilizing standard strain of S. aureus MTCC6625 was inoculated intravaginally at 10(8)cfu for 10 days followed by mating on day 12, fertility was observed in all the female mice. This supports the hypothesis that infertility observed in the former groups was as a result of colonization with spermagglutinating strain of S. aureus.