Oestradiol-induced infection of the genital tract of female mice by Mycoplasma hominis

Treatment of female BALB/c mice with oestradiol rendered them susceptible to vaginal colonization by three of four different strains of Mycoplasma hominis. Overall, the organisms were recovered persistently from the vagina of 68 (87%) of 78 of these mice. Strain TO mice given one of the strains were at least susceptible, all of ten becoming colonized and larger numbers of organisms being recovered. The hormone arrested the reproductive cycle in the oestrous phase, characterized by non-nucleated, cornified vaginal epithelial cells. In contrast, M. hominis organisms were isolated transiently from only seven (10.5%) of 66 BALB/c mice not treated with oestradiol, after intravaginal inoculation; treatment with progesterone, which induced the dioestrous phase of the cycle, did not render any of 10 BALB/c mice susceptible to vaginal colonization. The minimum number of organisms (2.5 x 10(5)) of one strain of M. hominis and the minimum dose of oestradiol (0.05 mg) required to induce persistent colonization were established. Vaginal colonization persisted for more than 200 d in some mice, the numbers of organisms recovered ranging between 10(1) and 10(8). At autopsy there was evidence of spread to the uterine horns and ovaries, and also to the oropharynx, of some animals but not to other organs. Infection was not associated with a polymorphonuclear leucocyte response in the vagina or elsewhere, but a fourfold serum antibody response to M. hominis, measured by the metabolism-inhibition technique, was detected in almost half of the mice tested.     

Inhibition of bacterial peptide deformylase by biaryl acid analogs

Peptide deformylase is an essential eubacterial metalloenzyme involved in the maturation of proteins by cleaving the N-formyl group from N-blocked methionine polypeptides. Biaryl acid analogs containing tetrazole, acyl sulfonamide, or carboxylate pharmacophores were found to be potent inhibitors of recombinant Escherichia coli peptide deformylase. Two of these compounds, a biphenyl tetrazole, compound 1, and a biphenyl acyl sulfonamide, compound 4, were competitive inhibitors with K(i) values of 1.2 and 6.0 microM, respectively. By analogy to the binding of related compounds to other metalloenzymes such as Bacteroides fragilis metallo-beta-lactamase CcrA and human carbonic anhydrase, a mechanism of inhibition is proposed for these peptide deformylase inhibitors where the acidic moieties form direct ionic interactions with the active site metal cation.     

Experimentally induced mycoplasmal infection in the genital tract of the female dog

After intrauterine inoculation of Mycoplasma canis in female dogs, the influence on uterus and general state of health was investigated using clinical, gynecological, microbiological and pathohistological methods.In three animals an enlargement of the uterus could be found at post mortem. These findings were identified as endometritis purulenta, intramural phlegmon and glandular cystic reaction of the endometrium.The microbiological and pathohistological findings are discussed in the light of the incidence of clinical symptoms and the influence of a mycoplasmal infection.     

Macrolactin N, a new peptide deformylase inhibitor produced by Bacillus subtilis

A new 24-membered ring lactone, macrolactin N, was isolated from a culture broth of Bacillus subtilis and its structure was established by various spectral analysis. Macrolactin N inhibited Staphylococcus aureus peptide deformylase with an IC50 value of 7.5 microM and also showed antibacterial activity against Escherichia coli and S. aureus.     

Immunization with chlamydial plasmid protein pORF5 DNA vaccine induces protective immunity against genital chlamydial infection in mice

To validate the immune protective efficacy of pORF5 DNA vaccine and to analyze potential mechanisms related to this protection. In this study, pORF5 DNA vaccine was constructed and evaluated for its protective immunity in a mouse model of genital chlamydial infection. Groups of BALB/c mice were immunized intranasally with pORF5 DNA vaccine. Humoral and cell mediated immune responses were evaluated. The clearance ability of chlamydial challenge from the genital tract and the chlamy- dia-induced upper genital tract gross pathology and histopathological characterization were also de- tected. The results showed that the total and the IgG2a anti-pORF5 antibody levels in serum were sig- nificantly elevated after pcDNA3.1-pORF5 vaccination, as were the total antibody and IgA levels in vaginal fluids. pcDNA3.1-pORF5 induced a significantly high level of Th1 response as measured by robust gamma interferon (IFN-γ). Minimal IL-4 was produced by immune T cells in response to the re-stimulation with pORF5 protein or the inactive elementary body in vitro. pcDNA3.1-pORF5-vacci- nated mice displayed significantly reduced bacterial shedding upon a chlamydial challenge and an accelerated resolution of infection. 100% of pcDNA3.1-pORF5 vaccinated mice successfully resolved the infection by day 24. pcDNA3.1-pORF5-immunized mice also exhibited protection against patho- logical consequences of chlamydial infection. The stimulated index was significantly higher than that of mice immunized with pcDNA3.1 and PBS (P<0.05). Together, these results demonstrated that immu- nization with pORF5 DNA vaccine is a promising approach for eliciting a protective immunity against a genital chlamydial challenge.     

Immunization with chlamydial plasmid protein pORF5 DNA vaccine induces protective immunity against genital chlamydial infection in mice

To validate the immune protective efficacy of pORF5 DNA vaccine and to analyze potential mechanisms related to this protection. In this study, pORF5 DNA vaccine was constructed and evaluated for its protective immunity in a mouse model of genital chlamydial infection. Groups of BALB/c mice were immunized intranasally with pORF5 DNA vaccine. Humoral and cell mediated immune responses were evaluated. The clearance ability of chlamydial challenge from the genital tract and the chlamydia-induced upper genital tract gross pathology and histopathological characterization were also detected. The results showed that the total and the IgG2a anti-pORF5 antibody levels in serum were significantly elevated after pcDNA3.1-pORF5 vaccination, as were the total antibody and IgA levels in vaginal fluids. pcDNA3.1-pORF5 induced a significantly high level of Th1 response as measured by robust gamma interferon (IFN-γ). Minimal IL-4 was produced by immune T cells in response to the re-stimulation with pORF5 protein or the inactive elementary body in vitro. pcDNA3.1-pORF5-vaccinated mice displayed significantly reduced bacterial shedding upon a chlamydial challenge and an accelerated resolution of infection. 100% of pcDNA3.1-pORF5 vaccinated mice successfully resolved the infection by day 24. pcDNA3.1-pORF5-immunized mice also exhibited protection against pathological consequences of chlamydial infection. The stimulated index was significantly higher than that of mice immunized with pcDNA3.1 and PBS (P<0.05). Together, these results demonstrated that immunization with pORF5 DNA vaccine is a promising approach for eliciting a protective immunity against a genital chlamydial challenge.     

1724例女性生殖道支原体感染调查及耐药性分析

目的了解本地区女性生殖道支原体的感染现状及耐药情况,为临床用药提供指导。方法采用生殖道支原体检测试剂盒,对2012年8月至2013年4月解放军第44医院妇科门诊就诊的1 724例女性生殖道标本进行支原体培养及药敏检测,并对结果做统计学分析。结果 1 724例女性生殖道支原体感染率为43.6%,其中单纯Uu感染占85.1%;单纯Mh感染占3.7%;Uu合并Mh感染占11.2%。感染人群中21-40岁年龄段占85.2%。支原体对交沙霉素、强力霉素、美满霉素耐药率较低,对环丙沙星、左氧氟沙星、红霉素及司帕沙星的耐药率较高,单纯Mh及Uu合并Mh感染对抗菌药物的耐药率高于单纯Uu感染。结论女性生殖道支原体感染以单纯Uu为主,感染人群多见于21-40岁,支原体对常用抗菌药物出现了不同程度的耐药性,建议临床根据药敏结果合理用药,以减少耐药菌株的产生。     

抵抗女性生殖道沙眼衣原体感染的免疫新策略

沙眼衣原体是引起沙眼和泌尿生殖道感染的主要病原体。据世界卫生组织2015年统计,全球每年约有1.3亿沙眼衣原体感染新发病例。研究表明CD4^+Th1型细胞免疫应答在抵抗沙眼衣原体感染中发挥着重要作用。因此,研究者依照抗沙眼衣原体感染的免疫应答特点,构建出许多候选疫苗,但都没有成功地应用于临床。近年研究发现,生殖道黏膜组织不仅存在体液免疫和细胞免疫,还驻留着一些引人注目的免疫细胞,提示增强黏膜免疫可作为预防沙眼衣原体感染的潜在途径,是抵抗生殖道沙眼衣原体感染的免疫新策略。本文全面概述了黏膜免疫与女性生殖道沙眼衣原体感染的研究进展,并为今后研制沙眼衣原体疫苗提供一些建议。     

女性生殖道支原体感染现状及耐药性调查分析

目的 了解女性生殖道支原体感染现状及耐药性,采取有效措施降低感染率,同时为临床合理用药提供依据.方法 对415例疑似生殖道感染标本采用珠海迪尔生物有限公司生产的试剂盒进行支原体培养和药敏试验.结果 415例女性生殖道标本中,支原体感染209例,阳性率为50.4％(209/415),其中单纯解脲脲原体(Uu)感染141例,阳性率34.0％(141/415),单纯人型支原体(Mh)感染25例,阳性率6.0％ (25/415),Uu和Mh混合感染43例,阳性率为10.4％ (43/415);药敏试验结果表明:Uu、Uu+ Mh、Mh对强力霉素、交沙霉素、美满霉素、四环素敏感性普遍较高,相反对红霉素、环丙沙星、罗红霉素、氧氟沙星等敏感性较差.结论 女性生殖道支原体感染率较高,已婚育龄妇女应加强妇科检查,做到早检查、早诊断、早治疗.同时,临床医生应结合药敏试验结果合理用药,以降低女性生殖道支原体感染的发生率.     

Peptide-immunoreactive nerves in the mammalian female genital tract

Summary Vasoactive intestinal polypeptide, substance P, neuropeptide Y and peptide histidine isoleucine immunoreactivities have been demonstrated in the female genitalia of rat, cat, mouse and guinea-pig using immunocytochemistry and radioimmunoassay. They were localized to nerves. Each type of immunoreactive nerve showed a distinct pattern of distribution, though all were associated to some degree with blood vessels and smooth muscle. Vasoactive intestinal polypeptide-immunoreactive and neuropeptide Y-immunoreactive nerves were the most abundant. Higher concentrations of peptides were detected in the female genitalia of the mouse than those of the other species studied. Vasoactive intestinal polypeptide-immunoreactive nerves were particularly concentrated in the cervix (89.1±17.2 pmol/g, mean±S.E.M.) and the uterus (57.4±14.8 pmol/g) of the mouse, while neuropeptide Y immunoreactivity was more abundant in the Fallopian tube of the mouse (31.6±11.8 pmol/g) and the vagina of the rat (38.6±4.8 pmol/g) than in other regions. Separate populations of ganglion cells in the paracervical ganglia were found to contain vasoactive intestinal polypeptide and neuropeptide Y immunoreactivities. Peptide histidine isoleucine-immunoreactive and vasoactive intestinal polypeptide-immunoreactive nerves were similarly distributed, but the former were much less frequent. Substance P-immunoreactive nerves were seen mainly beneath the epithelium of the vagina and were, in general, more numerous in the guinea-pig than in other species. The significance of these peptide-immunoreactive nerves in the female genital organ remains to be determined.Dr. Wang is on leave from The Institute of Acupuncture, The Academy of Chinese Traditional Medicine, Peking, China.     

Solvent-assisted slow conversion of a dithiazole derivative produces a competitive inhibitor of peptide deformylase

Due to its potential as an antibiotic target, E. coli peptide deformylase (PDF_Ec) serves as a model enzyme system for inhibitor design. While investigating the structural–functional and inhibitory features of this enzyme, we unexpectedly discovered that 2-amino-5-mercapto-1,3,4-thiadiazole (AMT) served as a slow-binding inhibitor of PDF_Ec when the above compound was dissolved only in dimethylformamide (DMF), but not in any other solvent, and allowed to age. The time dependent inhibitory potency of the DMF-dissolved AMT was correlated with the broadening of the inhibitor's 295 nm spectral band toward the visible region, concomitant with the increase in the mass of the parent compound by about 2-fold. These data led to the suggestion that DMF facilitated the slow dimerization of AMT (via the formation of a disulfide bond), and that the dimeric form of AMT served as an inhibitor for PDF_Ec. The latter is not caused by the simple oxidation of sulfhydryl groups by oxidizing agents such as H₂O₂. Newly synthesized dimeric/dithiolated form of AMT (“bis-AMT”) exhibited similar spectral and inhibitory features as given by the parent compound when incubated with DMF. The computer graphic modeling data revealed that bis-AMT could be reliably accommodated within the active site pocket of PDF_Ec, and the above enzyme–ligand interaction involves coordination with the enzyme resident Ni²⁺ cofactor. The mechanism of the DMF-assisted activation of AMT (generating bis-AMT), the overall microscopic pathway for the slow-binding inhibition of PDF_Ec by bis-AMT, and the potential of bis-AMT to serve as a new class of antibiotic agent are presented.     

Pre-natal innervation of the human female genital tract

In the human fetus of 14 weeks, ganglia on either sides of the Müllerian uterovaginal canal contained two types of cells. In the 16th week, axons invaded the basal zone of the stratified squamous epithelium at the sides of the upper vagina. In the 20th week, vesicular nuclei typified the large neurons in the midportion of the cervico-vaginal ganglion. During the 22nd week, capsulated ganglia invaded the wall of the upper vagina forming three concentrically disposed strata. Non-capsulated clusters invaded its lamina propria. At the 24th week, axons were shaded after reaching the superficial zone of the stratified vaginal epithelium. In the 28th week, satellites surrounded the mature neurons and sheath cells enveloped the axons. Ganglia invaded the splitted muscle layer of the upper vagina at 30 weeks. Intraepithelial fibres invaded the whole thickness of the endometrium, the columnar epithelium of the cervix and uterine tube at 40 weeks. Nerve cells were detected among the basal epithelial cells of the lower vagina and its subepithelial plexus.     

Amylase in human lungs and the female genital tract

Summary We investigated the localization of amylase in normal human lungs and the female genital tract using immunohistochemical and histochemical methods. Immunohistochemical procedures were applied to formaldehydefixed, paraffin-embedded specimens as well as to cryostat sections of periodate/lysine/paraformaldehyde (PLP)-fixed tissues. The starch-substrate-film method was used for the histochemical investigation of unfixed frozen sections. Amylase immunoreactivity was observed in ciliated epithelial cells of the bronchus and in serous cells of the bronchial glands but not in the alveolar epithelium. Immunoreactive amylase was also found in the cytoplasm of the ciliated epithelium of the fallopian tubes, especially in the apical part of the cytoplasm and in ciliary vesicles. Immunoreactive amylase was also found to be present in the surface epithelial cells and glands of the uterine cervix, as well as in the superficial part of the endometrial glands. The distribution of amylase activity revealed using histochemistry was similar to that observed in cryostat sections of PLP-fixed tissues after immunohistochemical staining. Amylase amtigenicity was better preserved in cryostat sections of PEP-fixed materials than in formaldehyde-fixed, paraffinembedded specimens. The results are discussed in relation to pulmonary and female-genital-tract diseases.This work was supported by a grant from the Tokyo Metropolitan Government     

Immunological responses to semen in the female genital tract

When spermatozoa, seminal plasma and semen extender reach the uterus and interact with local leukocytes and endometrial cells, several immune mechanisms are initiated which have immediate, mid-term and long-term effects on ovulation, sperm cell selection, fertilization and pregnancy success by assuring the acceptance of fetal tissues. This report gives an overview on relevant key immune mechanisms following roughly the time axis after insemination. Detailed knowledge regarding these mechanisms will aid maximizing reproductive efficiency in livestock production. In the future, the many species involved will require a more comparative approach, since evidence is growing that endometrial physiology and the response to varying amounts and compositions of seminal plasma, various semen extenders, and variable numbers of spermatozoa also provoke different immune responses.