The adhesion protein of Mycoplasma genitalium inhibits urethral epithelial cell apoptosis through CypA-CD147 activating PI3K/ Akt/NF-κB pathway.

By interacting with the receptor on the host cells membrane, Mycoplasma genitalium, a prokaryotic bacterium primarily transmitted through sexual contact, can adhere to and even enter cells. The adhesion protein of M. genitalium (MgPa) plays a critical function in the adhering and subsequent invasion into host cells. Our prior studies verified that cyclophilin A (CypA) was the receptor of MgPa on human urethral epithelial cells (SV-HUC-1) membrane and could induce pro-inflammatory cytokines production through the CypA-CD147-ERK-NF-κB pathway. This research aims to understand how MgPa interacts with its membrane receptor CypA to cause apoptosis in host cells. We employed flow cytometry to see if MgPa prevents or enhances apoptosis of SV-HUC-1 cells. The apoptosis-related proteins such as Bax, caspase-3, and cleaved caspase-3 were assayed using Western blot. Results suggested that MgPa could inhibit the apoptosis of SV-HUC-1 cells. And we demonstrated that interference with the expression of CypA or CD147 significantly reversed the inhibitory effect of MgPa on SV-HUC-1 cells apoptosis, indicating that MgPa inhibited urothelial cells apoptosis through CypA/CD147. Furthermore, we discovered that MgPa regulates the PI3K/Akt/NF-κB pathway through CypA/CD147 to inhibit SV-HUC-1 cells apoptosis. Ultimately, the inhibitory effect of MgPa on the apoptosis of the urothelial epithelial cells extracted from CypA-knockout mice was validated by Annexin V/PI assay. The results corroborated that MgPa could also inhibit mouse urothelial epithelial cells apoptosis. In summary, we demonstrated that MgPa could inhibit SV-HUC-1 cells apoptosis via regulating the PI3K/Akt/NF-κB pathway through CypA/CD147, providing experimental evidence for elucidating the survival strategies of M. genitalium in host cells. KEY POINTS: • M. genitalium protein of adhesion inhibited human urethral epithelial cells apoptosis through CypA-CD147 activating the signal pathway of PI3K/Akt/NF-κB • The knockdown of CypA and CD147 could downregulate the M. genitalium -activated PI3K/Akt/NF-κB pathway in SV-HUC-1 cells • MgPa could inhibit the apoptosis of normal C57BL mouse primary urethral epithelial cells, but not for CypA-knockout C57BL mouse primary urethral epithelial cells.

Molecular Advances on Phytases in Barley and Wheat.

Phytases are pro-nutritional enzymes that hydrolyze phytate and make associated nutrients, such as phosphorous, iron, and zinc, bioavailable. Single-stomached animals and humans depend on phytase supplied through the diet or the action of phytase on the food before ingestion. As a result, phytases-or lack thereof-have a profound impact on agricultural ecosystems, resource management, animal health, and public health. Wheat, barley and their Triticeae relatives make exceptionally good natural sources of phytase. This review highlights advances in the understanding of the molecular basis of the phytase activity in wheat and barley, which has taken place over the past decade. It is shown how the phytase activity in the mature grains of wheat and barley can be ascribed to the PAPhy_a gene, which exists as a single gene in barley and in two or three homeologous copies in tetra- and hexaploid wheat, respectively. It is discussed how understanding the function and regulation of PAPhy_a may support the development of improved wheat and barley with even higher phytase activity.

Cyclophilin A is the potential receptor of the Mycoplasma genitalium adhesion protein.

The Mycoplasma genitalium adhesion protein (MgPa), the most important outer membrane protein of M. genitalium, plays a vital role in the adhesion to and invasion of host cells by M. genitalium. Identification of MgPa receptors will help elucidate the pathogenic mechanism of M. genitalium. However, the receptor protein of MgPa has not been reported to date. In this study, an MgPa-binding protein with a molecular weight of approximately 17 kDa was screened from SV-HUC-1 cell membrane proteins by a modified virus overlay protein binding assay (VOPBA). Liquid chromatography-mass spectrometry (LC-MS) was used to analyze the protein components of the 17-kDa protein. The results demonstrated that the MgPa-binding protein was most likely Cyclophilin A (CyPA). The binding activity and distribution of CyPA in SV-HUC-1 cells were detected using indirect ELISA, western blotting, far-western blotting and indirect immunofluorescence. We found that recombinant MgPa (rMgPa) could bind with CyPA from SV-HUC-1 cell membrane proteins and to recombinant CyPA, which indicated that CyPA was predominant component of the 17-kDa protein band and can interact with rMgPa. In addition, an indirect immunofluorescence assay showed that CyPA was partially distributed on the membrane surfaces of SV-HUC-1 cells and could partially inhibit the adhesion of rMgPa and M. genitalium to SV-HUC-1 cells. Co-localization assays further indicated that rMgPa and M. genitalium can interact with CyPA. These results suggested that the CyPA located on SV-HUC-1 cell membranes may be the potential receptor of MgPa, which could provide an experimental basis for elucidating the function of MgPa and the possible pathogenic mechanism of M. genitalium.

Three polypeptides screened from phage display random peptide library may be the receptor polypeptide of Mycoplasma genitalium adhesion protein.

Mycoplasma genitalium adhesion protein (MgPa) is a major adhesin of M. genitalium, a human pathogen associated with a series of genitourinary tract diseases. MgPa plays a very important role in M. genitalium adhering to the host cells. However, the exact receptor peptides or proteins of MgPa are still poorly understood so far. Three polypeptides (V-H-W-D-F-R-Q-W-W-Q-P-S), (D-W-S-S-W-V -Y-R-D-P-Q-T) and (H-Y-I-D-F-R-W) were previously screened from a phage display random peptide library using recombinant MgPa (rMgPa) as a target molecule. In this study, three polypeptides were artificially synthesized and investigated as to whether they are potential receptors of MgPa. We found that rMgPa specifically bound to three synthesized polypeptides as determined via an indirect enzyme-linked immunosorbent assay (ELISA). Moreover, three polypeptides were further identified by indirect immunofluorescence microscopy (IFM). We confirmed that rMgPa and M. genitalium can adhere to SV-HUC-1 cells in vitro and that anti-rMgPa antibody and three synthesized polypeptides can partially inhibit the adherence of rMgPa and M. genitalium to SV-HUC-1 cells. In summary, these three polypeptides may be the essential receptor peptides of MgPa, and may aid in enhancing the understanding of biological function of MgPa and the possible pathogenic mechanism of M. genitalium.

The Relationship between Body Composition and Physical Fitness and the Effect of Exercise According to the Level of Childhood Obesity Using the MGPA Model.

Childhood obesity can lead to adulthood obesity with adverse effects. Since body composition and physical fitness differ depending on the obesity degree, a systemic analysis could help classify that degree. We used three study designs based on the obesity degree (body mass index [BMI] as a reference) for our objectives. First, we identified the relationship between body composition and physical fitness. Second, we determined the effects of exercise on body composition and physical fitness. Third, we performed a path analysis of the impact of exercise on body composition and physical fitness, and verified those effects among the groups. In study 1, 164 10-year-old subjects were divided into four groups: 33 in the normal weight (NO), 34 in overweight (OV), 54 in obesity (OB), and 43 in the severe obesity (SOB) group. In study 2, 101 participants from study 1 who wished to participate in the exercise program were divided into four groups (same criteria). The exercise program (three times a week for 60 min, for 16 weeks) consisted of sports and reinforcement exercises of increasing intensity. Body composition was measured by body weight, percentage of body fat (%BF), muscle mass, skeletal muscle mass (SMM), and body mass index (BMI). In contrast, physical fitness was measured by muscular strength, flexibility, muscular endurance, agility, and balance. As a result, all body composition variables were higher in the SOB group than in the other groups. Physical fitness, muscular strength and balance, and agility were highest in the SOB, NO, and OV groups, respectively. Pearson's correlation revealed that muscular strength was associated with height and body weight across all groups. Agility showed a negative correlation with %BF in the NO, OB, and SOB groups. SMM was positively correlated in the OB and SOB groups. After the exercise intervention, BMI and the %BF of the SOB group were significantly reduced (p < 0.01, and p < 0.001, respectively), while SMM presented a significant increase (p < 0.001). Height also showed a significant increase in all groups (p < 0.001). Among physical fitness variables, muscular strength, flexibility, muscular endurance, and balance showed a significant increase in all groups, while a significant increase in power was observed in only the OB and SOB groups. As for the effects of the body composition on physical fitness after exercise intervention, the greatest impact was observed for balance, muscular strength and agility, and muscular endurance in NO, OV, and OB groups, respectively. In conclusion, the body composition, physical fitness relationship, and the effects of exercise intervention on them differed depending on the obesity degree. Furthermore, the results varied according to the obesity degree. Thus, our study highlights the importance of creating particular exercise programs for the effective prevention and treatment of childhood obesity considering the obesity degree.

Identification of histone H2B as a potential receptor for Mycoplasma genitalium protein of adhesion.

Mycoplasma genitalium, the smallest prokaryotic microorganism capable of independent replication, is increasingly recognized as a sexually transmitted pathogen. M. genitalium protein of adhesion (MgPa) plays a pivotal role in the process of M. genitalium adhesion to host cells. We previously identified cyclophilin A as a cellular receptor of MgPa using the virus overlay protein binding assay (VOPBA) together with liquid chromatography-mass spectrometry (LC-MS). In the current study, we have evaluated H2B as an alternative cellular receptor for MgPa since H2B was assigned the second higher score as a potential binding partner of MgPa in the VOPBA and LC-MS screen. It was found that recombinant MgPa specifically bind to H2B both in the SV-HUC-1 cell membrane and in form of a recombinant protein. H2B was detected throughout the SV-HUC-1 cells, including the cytoplasmic membrane, cytosol and nucleus. Importantly, H2B partially inhibited the adhesion of M. genitalium to SV-HUC-1 cells. Finally, H2B was both co-precipitated with recombinant MgPa and co-localized with M. genitalium and recombinant MgPa in SV-HUC-1 cells. The above observations suggest that H2B may act as a potential cellular receptor of MgPa for mediating M. genitalium adhesion to host cells.

Mycoplasma genitalium Protein of Adhesion Promotes the Early Proliferation of Human Urothelial Cells by Interacting with RPL35.

Mycoplasma genitalium is a newly recognized pathogen associated with sexually transmitted diseases (STDs). MgPa, the adhesion protein of Mycoplasma genitalium, is the main adhesin and the key factor for M. genitalium interacting with host cells. Currently, the long-term survival mechanism of M. genitalium in the host is not clear. In this study, a T7 phage-displayed human urothelial cell (SV-HUC-1) cDNA library was constructed, and the interaction of MgPa was screened from this library using the recombinant MgPa (rMgPa) as a target molecule. We verified that 60S ribosomal protein L35 (RPL35) can interact with MgPa using far-Western blot and co-localization analysis. According to the results of tandem mass tag (TMT) labeling and proteome quantitative analysis, there were altogether 407 differentially expressed proteins between the pcDNA3.1(+)/MgPa-transfected cells and non-transfected cells, of which there were 6 downregulated proteins and 401 upregulated proteins. The results of qRT-PCR demonstrated that interaction between rMgPa and RPL35 could promote the expressions of EIF2, SRP68, SERBP1, RPL35A, EGF, and TGF-ß. 3-(4,5)-Dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide bromide (MTT) assays corroborated that the interaction between rMgPa and RPL35 could promote SV-HUC-1 cell proliferation. Therefore, our findings indicated that the interaction between rMgPa and RPL35 can enhance the expressions of transcription-initiation and translation-related proteins and thus promote cell proliferation. This study elucidates a new biological function of MgPa and can explain this new mechanism of M. genitalium in the host.

MgpB Types among Mycoplasma genitalium Strains from Men Who Have Sex with Men in Berlin, Germany, 2016-2018.

Mycoplasma genitalium is a cell wall-less bacterium causing urethritis and other sexually transmitted diseases. Despite a strongly conserved genome, strains in clinical samples can be typed by different methods. To obtain data from the risk population of men having sex with men, we analyzed the typing region in the gene coding for the MgpB adhesin of M. genitalium first in 163 and 45 follow-up samples among patients of two specialized practices in Berlin, Germany. Strains belong to 43 different mgpB types emphasizing the diversity of the genome region. With respect to 133 types previously described, 27 new types were found. However, the majority of strains (64.4%) were assigned to types 4, 6, 113, and 108, respectively. A correlation between mgpB type and the occurrence of mutations associated with macrolide and quinolone resistance was not demonstrated. Investigation of follow-up samples from 35 patients confirmed the same mgpB and, additionally, MG_309 types in 25 cases. In 10 cases, differences between types in subsequent samples indicated an infection with a genetically different strain in the period between samplings. MgpB/MG_309 typing is a useful method to compare M. genitalium strains in samples of individual patients as well as those circulating in different populations.

Identification of protein that interacts with adhesion protein of Mycoplasma genitalium from T7-phage display cDNA library / 中国免疫学杂志

Objective:To screen and identify the protein that interacts with the adhesion protein of Mycoplasma genitalium (MgPa)from T7-phage display cDNA library of human uroepithelial cells(SV-HUC-1).Methods:Recombinant adhesion protein of My-coplasma genitalium(rMgPa)was used as target molecule to biopan the T7 phage display cDNA library of SV-HUC-1 cell,the selected positive clones were analysed using DNA sequencing and BLAST analysis and identified by means of indirect ELISA,Dot immunoblot and Far-western blot.Results:After four rounds of biopanning,positive phages were obviously enriched.According to the results of DNA sequencing and BLAST analysis,the selected randomly 32 positive clones included 7 kinds different sequences,of which the number of RPL35 repeats was the most.The results of indirect ELISA,Dot immunoblot and Far-western blot showed that 7 representative phages could bind specifically with rMgPa.Conclusion:60S ribosomal protein L35(RPL35) may be the interacting protein of MgPa,which lays the experimental foundation for understanding the function of MgPa and the pathogenesis of Mycoplasma genitalium.

Detección de Mycoplasma genitalium y correlación con manifestaciones clínicas en una población del estado Zulia, Venezuela / Mycoplasma genitalium detection and correlation with clinical manifestations in population of the Zulia State, Venezuela

Diverse studies demonstrate an association between Mycoplasma genitalium and urogenital pathologies. The aim of this study was to investigate the prevalence of M. genitalium in patients attending gynecological evaluation in private clinics (n = 172). DNA amplification assays of the genes 16S rRNA and MgPa were utilized. The prevalence of M. genitalium in the study population was 7.5 percent. M. genitalium was detected in 12.1 percent and 4.1 percent of the symptomatic and asymptomatic patients, respectively (p = 0.047). The infection was diagnosed in patients with cervicitis (17.2 percent) and mucopurulent secretion (16.6 percent) and the highest prevalence of infections was registered in the 31-40 years age group. No significant association between the presence of M.genitalium and individual clinical manifestations or the patients age was showed (p > 0.05). The high prevalence of M. genitalium infections, mostly in patients with clinical manifestations showed in this study, warrants the application of diagnostic strategies in the population to investigate the clinical meaning of these microorganisms and to reevaluate therapeutic schemes against non-gonococcal and non-chlamydial infections.

Diversos estudios demuestran una asociación entre Mycoplasma genitalium y patologías urogenitales. El objetivo de este trabajo fue investigar la prevalencia de infecciones por M. genitalium en pacientes atendidas en clínicas privadas (n = 172). Se utilizaron ensayos de amplificación de genes 16S rARN y MgPa. La prevalencia de M. genitalium en esta población fue 7,5 por ciento. Mycoplasma genitalium fue detectado en 12,1 y 4,1 por ciento) de las pacientes sintomáticas y asintomáticas, respectivamente (p = 0,047). La infección se diagnosticó en pacientes con cervicitis (17,2 por ciento) y con secreción mucopurulenta (16,6 por ciento) y la mayor prevalencia de infecciones se registró en el grupo etario de 31 a 40 años. No se encontró asociación significativa entre la presencia de M. genitalium y manifestaciones clínicas individuales o edad de las pacientes (p > 0,05). La alta prevalencia de infecciones por M. genitalium, principalmente en pacientes con manifestaciones clínicas demostrada en este estudio, demanda la aplicación de estrategias diagnósticas en la población para investigar el significado clínico de estos microorganismos y reevaluar esquemas terapéuticos contra infecciones no gonocóccicas y no clamidiales.