Ajonuma L C
Ajonuma L C
Publications Authored By Ajonuma L C
Eighty patients undergoing hemodialysis in the dialysis unit of our hospital were enrolled for this study. Sixty-nine patients completed the study. Comparative analysis revealed significant difference in social isolation with favorable result for the Filipino patients. Other measures correlate well although with differences that were not statistically significant. NHP can be successfully applied as a standard QOL tool in the Philippines. However, it should be translated into Filipino to avoid language difficulty. NHP may be recommended for QOL determination in other developing countries.
pneumoniae infection in the pathogenesis of periodontal disease remains unknown. The present hypothesis proposes that C. pneumoniae is involved in the pathogenesis of periodontal diseases. This will lead to a better understanding of the etiopathogenesis of periodontal disease, better treatment strategy and savings on total health care costs.
Here we report that CFTR (cystic fibrosis transmembrane conductance regulator), an apical epithelial anion channel, is required for cellular entry and internalization of C. trachomatis. Human epithelial cell lines expressing functional CFTR internalized more C. trachomatis than the cells expressing mutant Delta508 CFTR. The in vitro cellular uptake of C. trachomatis can be blocked by CFTR inhibitors or antibody, and the in vivo cellular uptake of C. trachomatis in CFTR mutant (CFTR(-/-)) mice was significantly less compared with that in the wild-type. Direct interaction between CFTR and C. trachomatis LPS (lipopolysaccharide) is demonstrated by their immune-co-localization and co-immunoprecipitation. Despite an increase in CFTR expression observed upon C. trachomatis LPS challenge, a reduction in its ion channel activity is observed, consistent with the notion that CFTR functions as a receptor for cellular entry and internationization of C. trachomatis, with compromised ion-channel function. These findings, for the first time, demonstrate that CFTR functions as a cell-surface receptor for epithelial cell entry, and internalization of C. trachomatis and these findings may lead to the development of new treatment strategies to curtail the spread of chlamydial infections.
University laboratory animal service center.
Adult female mice with regular estrous cycles.
Intrauterine injection of C. trachomatis lipopolysaccharide (LPS), tumor necrosis factor-alpha (TNF-alpha), and estrogen (E) at diestrus and preimplantation.
The CFTR messenger RNA (mRNA) and protein levels were evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot, respectively, in mouse uterus treated with C. trachomatis LPS, TNF-alpha or E. Endometrial electrolyte transport and uterine fluid accumulation were determined by the short circuit current and uterine wet weight, respectively. Number of implanted embryos was also counted to demonstrate the effect of treatments.
Uterine C. trachomatis LPS infection induced up-regulation of CFTR expression with enhanced anion secretion, abnormal fluid accumulation in mouse uterus at diestrus, and reduced implantation rate. Administration of exogenous TNF-alpha to mouse uterus mimicked the C. trachomatis LPS infection-induced CFTR up-regulation, enhanced CFTR channel activity, and fluid accumulation. Abnormal uterine fluid accumulation and implantation failure were also observed when CFTR was up-regulated by E.
The present results suggest that C. trachomatis infection-induced release of cytokines could abnormally up-regulate CFTR expression leading to abnormal uterine fluid accumulation, which may result in infertility often associated with C. trachomatis infection.
We performed this study to determine the involvement of cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-activated chloride channel that regulates epithelial electrolyte and fluid secretion, in hydrosalpinx fluid formation.
Western blot analysis was used to determine CFTR expression in the hydrosalpinges that were seen on the ultrasound scans of infertile assisted reproduction treatment patients. Correlation with C. trachomatis infection was done by testing patients' sera for C. trachomatis immunoglobulin G antibody titer using a Capita enzyme-linked immunosorbent assay based kit. CFTR involvement was further verified in a rat C. trachomatis infection model and confirmed using CFTR mutant (CFTR(tm1Unc)) mice.
Here we report on the up-regulated expression of CFTR in the hydrosalpinx tissues of infertile patients with detectable serum levels of C. trachomatis antibody (immunoglobulin G). In a rat model, increased CFTR expression and fluid accumulation could be observed in the uterine horns infected with C. trachomatis elementary bodies, which was reversed by antibiotics treatment. In C. trachomatis-infected CFTR(tm1Unc) mice, however, no detectable fluid accumulation was observed.
These findings suggest the involvement of CFTR in the pathogenesis of hydrosalpinx fluid formation and may provide grounds for a better treatment strategy to improve assisted reproduction treatment outcome in infertile patients with hydrosalpinx.
However, the underlying mechanism for tissue fluid secretion induced by C. trachomatis and most of other infectious pathogens is not known. Here, we report that in mice C. trachomatis infection models, the expression of cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP activated chloride channel, is up regulated together with increased cytokine release and tissue fluid accumulation that can be reversed by treatment with antibiotic specific for C. trachomatis and CFTR channel blocker. However, C. trachomatis infection cannot induce tissue edema in CFTRtm1Unc mutant mice. Administration of exogenous IL-1beta to mice mimics the C. trachomatis infection-induced CFTR upregulation, enhanced CFTR channel activity and fluid accumulation, further confirming the involvement of CFTR in infection-induced tissue fluid secretion.
The present hypothesis aims to discuss the possible role of epithelial ion channels particularly cystic fibrosis transmembrane conductance regulator (CFTR) involvement in the pathogenesis of OHSS. This may provide grounds for the development of a better treatment strategy to reduce the risk of OHSS and improve in vitro fertilization (IVF) outcome.
Disturbance of the fluid microenvironment due to defects or abnormal regulation of these ion channels as causes for a number of pathological conditions, such as ovarian hyperstimulation syndromes, hydrosalpinx and infertility, is also discussed.
Upon ovarian hyperstimulation, rats develop OHSS symptoms, with up-regulated CFTR expression and enhanced CFTR channel activity, which can also be mimicked by administration of estrogen, but not progesterone, alone in ovariectomized rats. Administration of progesterone that suppresses CFTR expression or antiserum against CFTR to OHSS animals results in alleviation of the symptoms. Furthermore, ovarian hyperstimulation does not induce detectable OHSS symptoms in CFTR mutant mice. These findings confirm a critical role of CFTR in the pathogenesis of OHSS and may provide grounds for better assisted reproduction treatment strategy to reduce the risk of OHSS and improve in vitro fertilization outcome.
Hematoxylin and eosin staining of HSP showed areas without epithelial cell lining or with abnormalities such as flattening of the epithelial layer and exfoliation of epithelial cells with occasional normal columnar epithelial lining. HSP muscle fibers were atrophic and occasionally replaced by fibrous tissues, or separated by areas of severe edema. Inflammatory cells could be found in hydrosalpinx fluid (HF) in the lumen in areas with flattened to no epithelial cells, without epithelial lining, as well as in dilated blood vessels and/or lymph vessels. Scanning electron microscopy of the epithelial surface revealed epithelial denudation-severe loss of both cilia and microvilli and stomata exuding globular bodies on eroded ampulla surfaces. Severe chronic inflammation and damage to the epithelial lining and musculature of Fallopian tubes and the presence of inflammatory cells provides an explanation for HF formation, and thus for the detrimental effects of HF on reproductive processes and IVF outcome.
The sperm count and motility, in vivo and in vitro fertilization rate as indicated by two-cell embryos and blastocyst rate were examined. The sperm count and motility from all three knockout mice were not significantly different from that of the wild type. Inducible NOS (iNOS) knockout mice were found to have the largest number of two-cell embryos/mouse collected after fertilization in vivo (P<0.01), but the rate of blastocyst formation from two-cell embryos in vitro was similar for all three knockouts. The rate of in vitro fertilization using either iNOS-deficient sperm or oocytes, but not those deficient in the other two NOS isoforms, was significantly elevated when compared to that in the wild type (P<0.001). While all three types of NOS do not seem to play a significant role in pre-ejaculated sperm function, iNOS may play an inhibitory role in sperm and oocyte functions affecting the process of fertilization and early embryo development.
Masson's trichrome staining showed areas of epithelial transformation, focally attenuated and pseudostratified. Immunostaining showed enhanced CFTR immunoreactivity in the focally attenuated and pseudostratified areas of HSP epithelium. RT-PCR revealed that CFTR expression in HSP was significantly greater than that in normal Fallopian tubes.
These results indicate that HSP epithelium undergoes epithelial transformation with elevated CFTR expression, which may lead to increased transepithelial electrolyte and fluid secretion resulting in HF formation. The present findings may lead to the development of new treatment strategies for infertile patients with HSP.
Semi-quantitative reverse transcriptase polymerase chain reaction and immunohistochemistry were used to study the expression and localization of CFTR and ENaC in uteri collected from mature superovulated female mice. RT-PCR showed maximal ENaC and CFTR expression on day 3 after mating. Maximal immunoreactivity was also observed for both ENaC and CFTR on day 3 after mating. However, ENaC was immunolocalized to the apical membrane of both luminal and glandular epithelia, while CFTR was predominantly found in the stromal cells rather than the epithelial cells. Differential expression and localization of CFTR and ENaC provide a molecular mechanism by which maximal fluid absorption can be achieved immediately prior to implantation, to ensure the immobilization of the blastocyst necessary for implantation.
Co-culture of sperm with endometrial cells treated with antisense oligonucleotide against CFTR, or with bicarbonate secretion-defective CF epithelial cells, resulted in lower sperm capacitation and egg-fertilizing ability. These results are consistent with a critical role of CFTR in controlling uterine bicarbonate secretion and the fertilizing capacity of sperm, providing a link between defective CFTR and lower female fertility in CF.
The information obtained may be useful for designing future in vitro culture models to investigate the functional roles of these ion channels in the endometrium.
Normal Fallopian tubes (n = 6) and hydrosalpinges (n = 9) were used to prepare epithelial cell culture and CM. Epithelial cell characterization was confirmed using electron microscopy. Sperm motility and acrosome reaction were determined using computer-aided sperm analysis and acrobead assay respectively and embryo development by mouse embryo development assay.
The percentage of human motile sperm incubated in hydrosalpinx CM was significantly different from those in normal Fallopian tube (NFT) CM and modified human tubal fluid medium (hTF) (control) (P < 0.05 at 3 h and P < 0.001 at 5 and 24 h), with alteration in movement characteristic, linearity, 24 h after incubation in hydrosalpinx CM (P < 0.05). However, other sperm movement characteristics remained unchanged. Reduced acrosome reaction and poor mouse embryo development were also observed in hydrosalpinx CM but not in NFT CM and hTF.
The results suggest that hydrosalpinx epithelial cells may be producing a fluid milieu hostile to sperm and early embryo development. The established epithelial cell culture system may provide a model to further investigate the mechanisms underlying the toxic effects of HF on embryo development and the adverse effects on IVF outcomes.
This review discusses the mechanisms underlying hydrosalpinx formation and its adverse effect on IVF outcome, with new insights into possible involvement of Fallopian tube epithelial transporters and ion channels, particularly the cystic fibrosis transmembrane conductance regulator (CFTR). Possible links between Chlamydia trachomatis in pelvic inflammatory disease and the subsequent CFTR-mediated events in hydrosalpinx formation leading to infertility in hydrosalpinx are proposed. The causes of reduced implantation, particularly in patients with visible hydrosalpinges shown on ultrasound scanning, are re-examined in light of these possible mechanisms.
Fifteen infertile patients with hydrosalpinx shown on ultrasound monitoring during ovarian stimulation underwent aspiration of HF after egg collection. Electrolytes, glucose and pyruvate concentrations were within the physiological ranges found in normal human tubal fluid. Sperm motility and velocities remained unchanged after 5 h of incubation with various concentrations of HF but the percentage of motile spermatozoa was significantly reduced after 24 h of incubation. Both 50 and 100% HF were potentially cytotoxic (survival indices <85%). The detrimental effect seemed to be dependent on the concentrations of HF. Low osmolarity, low lactate concentrations or the protein content may be responsible for the loss of sperm motility. A human sperm survival test using HF may be useful in selecting appropriate treatment options for patients with hydrosalpinx undergoing IVF treatment or tubal surgery.