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Trichomonas vaginalis (T. vaginalis) is a widespread and important sexually transmitted pathogen. Adherence to the surface of the host cell is the precondition forthis parasite's parasitism and pathogenicity. Adhesion protein 65 (TvAP65) plays a key role in the process of adhesion. However, how TvAP65 mediates the adhesion and pathogenicity of T. vaginalis to host cellsis unclear. In this study, we knocked down the expression of TvAP65 in trophozoites by small RNA interference. The number of T. vaginalis trophozoites adhering to VK2/E6E7 cells was decreased significantly, and the inhibition of VK2/E6E7 cells proliferation and VK2/E6E7 cells apoptosis and death induced by T. vaginalis were reduced, after the expression of TvAP65 was knocked down. Animal challenge experiments showed that the pathogenicity of trophozoites was decreased by passive immunization with anti-rTvAP65 PcAbs or blocking the TvAP65 protein. Immunofluorescence analysis showed that TvAP65 could bind to VK2/E6E7 cells. In order to screen the molecules interacting with TvAP65 on the host cells, we successfully constructed the cDNA library of VK2/E6E7 cells, and thirteen protein molecules interacting with TvAP65 were screened by yeast two-hybrid system. The interaction between TvAP65 and BNIP3 was further confirmed by coimmunoprecipitation and colocalization. When both TvAP65 and BNIP3 were knocked down by small RNA interference, the number of T. vaginalis adhering to VK2/E6E7 cells and the inhibition of VK2/E6E7 cells proliferation were significantly lower than those of the group with knockdown of TvAP65 or BNIP3 alone. Therefore, the interaction of TvAP65 and BNIP3 in the pathogenesis of T. vaginalis infecting host cells is not unique and involves other molecules. Our study elucidated that the interaction between TvAP65 and BNIP3 mediated the adhesion and pathogenicity of T. vaginalis to host cells, provided a basis for searching for the drug targets of anti-T. vaginalis, and afforded new ideas for the prevention and treatment of trichomoniasis., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2023. Published by Elsevier B.V.)

Trichomonas vaginalis (T. vaginalis) infection is the most common non-viral sexually transmitted disease (STD) in the world. It can cause male reproductive dysfunction and infertility. However, the pathogenic mechanism is not clear. In this study, the excretory secretory proteins of T. vaginalis (TvESPs) were collected, concentrated, and sterilized. After sperm co-cultured with TvESPs, the survival rate and motility of sperms were analyzed by seminal routine examination, and the results showed that the TvESPs could significantly reduce the survival rate and motility of sperms. Fluorescence staining displayed that TvESPs could destroy the integrity of sperm acrosomes. Flow cytometry indicated that TvESPs induced sperm apoptosis. By mouse in vitro fertilization, we confirmed that TvESPs could significantly reduce the fertilization ability of sperms and negatively affect the development of the fertilized ovum. Via semi-quantitative analysis, we found that the apoptosis-related p27, SMAC, p53, BAX, BCL-2, XIAP, and BCL-W molecules were down-regulated in mouse sperm cells after interaction between the sperms and TvESPs, which played an important role in regulating sperm apoptosis. In conclusion, our study showed that T. vaginalis degraded semen quality and negatively affected male fertility by TvESPs. TvESPs may damage sperms by breaking the balance between sperm pro-apoptotic and anti-apoptotic molecules. This study proves that T. vaginalis infection is a risk factor for infertility., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2022. Published by Elsevier B.V.)

Trichomoniasis, an infectious disease caused by a parasite called Trichomonas vaginalis (T. vaginalis), enhances the risk of HIV infection, cervical and prostate cancer, and infertility. Therefore, efforts have to be made for accurate, specific, and rapid diagnosise and treatment of trichomoniasis. Today, optical nanosensors have created an opportunity for diagnosis without sophisticated and expensive tools and the need for expertise; at the same time, they are highly sensitive and fast. An optical nano-genosensor was designed by conjugation of gold nanoparticles and a specific oligonucleotide (AuNPs-probe) from repeated DNA target for specific and sensitive polymerase chain reaction diagnosis of T. vaginalis gene sequence (L23861.1). The hybridization of AuNPs-probe was investigated with different concentrations of complementary sequence in synthesized target, gene sequence of standard T. vaginalis genomic DNA extraction, and PCR products of genomic DNA samples extracted from patients. Negative samples including synthesized non-complementary sequence, genomics DNA of other pathogens, and genomics DNA of healthy persons were considered for proof of the accuracy of the sensor function. The occurrence of correct hybridization was detected by adding acid to the medium and observing the changes in the color of the medium and spectroscopic spectrum. Based on spectrophotometric results, the fabricated genosensor had detection limits of 35.16 and 31 pg μL -1 for the detection of synthetic target and genomic DNA sequences, respectively. The results confirmed the correct function of genosensor for the detection of T. vaginalis in clinical samples. Advantages such as low cost, visual detection, speed, and easy diagnosis encourage the use of this sensor in pathogen detection in the future., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Trichomonas vaginalis (T. vaginalis) is a common sexually transmitted infection, affecting the urogenital tract. Trichomoniasis is customarily treated with metronidazole (MTZ). MTZ is known to cause undesirable side effects and there is several reports on MTZ resistant T. vaginalis. Thus, the present study aimed to in-vitro evaluate the activity of DNA minor groove binder drug ''Netropsin dihydrochloride'' against metronidazole-sensitive T. vaginalis isolates (G and U isolates) and resistant T. vaginalis isolate (ATCC50138) (R isolate). Netropsin was tested at concentrations ranging from 3.5 to 200 μg/ml. It showed effectiveness against all isolates with MLC of 12.5 μg/ml for G and U isolates and of 25 μg/ml for R isolate. Cytotoxicity assay of isolates exposed to the respective MLC of netropsin for 42 h showed a highly significant reduction in the death percentage of MCDK cell line as compared to the effect elicited by drug free controls. The hemolytic activity was evaluated by hemolytic assay and by monitoring the interaction of T. vaginalis isolates with human erythrocytes by inverted microscopy and scanning electron microscopy. The hemolytic assay showed (0%) hemolysis of RBCs incubated with T. vaginalis isolates treated with the corresponding MLC of netropsin for 24 h. Scanning electron microscopy revealed cytoskeletal deformities of netropsin treated isolates. Taken together, these observations suggest that netropsin is a promising therapy for T. vaginalis infection affecting its viability, virulence, cytopathogenic and hemolytic activity with a mechanism of action that might overcome T. vaginalis resistance to metronidazole., (Copyright © 2019 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)