A reliable enzyme-linked immunosorbent assay for the determining of sesame proteins in raw food ingredients and in processed foods.

Sesame is a frequent cause of adverse food reactions in allergic patients. We developed a novel sandwich enzyme-linked immunosorbent assay (ELISA) using two monoclonal antibodies and a unique extraction buffer for the detection and quantification of sesame proteins in processed foods and in raw food ingredients to clarify the validity of sesame labeling and for precautionary allergen labeling. The developed sandwich ELISA method is highly specific for sesame proteins. The limit of detection (LOD) and limit of quantification (LOQ) are 0.013 µg/g and 0.025 µg/g, respectively. The recoveries for incurred food samples, such as dressing, breads, sauce and pudding, ranged from 67 % to 81 %, while the repeatability and reproducibility coefficients of variation were less than 4.7 % and 4.5 %, respectively. The developed method has applicability for food products and is a reliable tool for the detection of hidden sesame proteins in raw food ingredients and in processed foods.

Thermal Molding of Organic Thin-Film Transistor Arrays on Curved Surfaces.

In this work, a thermal molding technique is proposed for the fabrication of plastic electronics on curved surfaces, enabling the preparation of plastic films with freely designed shapes. The induced strain distribution observed in poly(ethylene naphthalate) films when planar sheets were deformed into hemispherical surfaces clearly indicated that natural thermal contraction played an important role in the formation of the curved surface. A fingertip-shaped organic thin-film transistor array molded from a real human finger was fabricated, and slight deformation induced by touching an object was detected from the drain current response. This type of device will lead to the development of robot fingers equipped with a sensitive tactile sense for precision work such as palpation or surgery.

Physical Property Evaluation of ZnO Thin Film Fabricated by Low-Temperature Process for Flexible Transparent TFT.

The usual silicon-based display back planes require fairly high process temperature and thus the development of a low temperature process is needed on flexible plastic substrates. A new type of flexible organic light emitting transistor (OLET) had been proposed and investigated in the previous work. By using ultraviolet/ozone (UV/O3) assisted thermal treatments on wet processed zinc oxide field effect transistor (ZnO-FET), through low-process temperature, ZnO-FETs were fabricated which succeeded to achieve target drain current value and mobility. In this study, physical property evaluation of ZnO was conducted in term of their crystallinity, the increase composition of ZnO formed inside the thin film and the decrease of the carbon impurities originated from aqueous solution of the ZnO itself. The X-ray diffraction (XRD) evaluation showed UV/03 assisted thermal treatment has no obvious effect towards crystallinity of ZnO in the range of low process temperature. Moreover, through X-ray photoelectron spectroscopy (XPS) evaluation and Fourier transform infrared (FT-IR) spectroscopy evaluation, more carbon impurities disappeared from the ZnO thin film and the increase of composition amount of ZnO, when the thin film was subjected to UV/O3 assisted thermal treatment. Therefore, UV/O3 assisted thermal treatment contributed in carbon impurities elimination and accelerate ZnO formation in ZnO thin film, which led to the improvement in the electrical property of ZnO-FET in the low-process temperature.

Partial Dissolution of Charge Order Phase Observed in ß-(BEDT-TTF)2PF6 Single Crystal Field Effect Transistor.

The effect of an applied gate electric field on the charge-order phase in ß-(BEDT-TTF)2PF6 single-crystal field-effect transistor structure was observed at around room temperature by technical improvement with respect to sample preparation and electrical measurements. A relatively slight but systematic increase of the electrical conductance induced by the applied gate electric field and its temperature dependence was observed at around the metal-insulator transition temperature (TMI). The temperature dependence of the modulated electrical conductance demonstrated that TMI was shifted toward the lower side by application of a gate electric field, which corresponds to partial dissolution of the charge-order phase. The thickness of the partially dissolved charge order region was estimated to be several score times larger than the charge accumulation region.

Protein kinase C (PKC) increases TACE/ADAM17 enzyme activity in porcine ovarian somatic cells, which is essential for granulosa cell luteinization and oocyte maturation.

During in vitro maturation of porcine cumulus cell-oocyte complexes and in vitro luteinization of porcine granulosa cells, FSH induces the expression of the protease TNFα-converting enzyme/A disintegrin and metalloproteinase domain 17 (TACE/ADAM17) and the epidermal growth factor (EGF)-like factors, which activate the EGF receptor (EGFR)-MAPK3/1 pathway in both cumulus and granulosa cells. FSH is known to activate not only protein kinase A and p38MAPK pathways in both cell types but also activates protein kinase C (PKC). Because PKC-induced association of cellular-Sarcoma (c-Src) and TACE/ADAM17 is required for TACE/ADAM17 enzyme activation in some cancer cells, we hypothesized that PKC and c-Src impact TACE/ADAM17-mediated activation of EGFR signaling pathway in porcine granulosa and cumulus cells. When granulosa cells or cumulus cell-oocyte complexes were cultured with FSH, PKC activity and c-Src phosphorylation increased and were associated with increased TACE/ADAM17 enzyme activity. The PKC inhibitor calphostin C (CalC) and the c-Src inhibitor (4 amino 5 (4 chlorophenyl) 7 (t butyl)pyrazolo[3,4 d]pyrimidine [PP2]) suppressed TACE/ADAM17 enzyme activity, whereas these inhibitors did not affect Tace/Adam17 mRNA expression. Immunoprecipitation analysis showed that FSH mediated the association of c-Src with TACE/ADAM17 via a PKC-dependent mechanism. Either CalC or PP2 suppressed EGFR downstream signaling pathway (MAPK3/1) in these ovarian cell types and reduced cumulus expansion, meiotic maturation of oocytes, and progesterone production. The negative effects were overcome by the addition of amphiregulin. Collectively, these results indicate that activation of TACE/ADAM17 via a PKC-induced c-Src-dependent manner mediates proteolytic activation of the EGF-like factors that are involved in the induction of granulosa cell differentiation, cumulus expansion, and meiotic maturation of porcine oocytes in vitro.

Granulysin produced by uterine natural killer cells induces apoptosis of extravillous trophoblasts in spontaneous abortion.

Immune changes are known to occur in recurrent spontaneous abortion, but it is unclear whether either maternal natural killer (NK) cells or T cells attack fetus-derived trophoblasts. To clarify the immunological causes of spontaneous abortion, we examined the relationship between cytotoxic granule proteins in decidual lymphocytes, such as granulysin, granzyme B, and perforin, and the induction of apoptosis in extravillous trophoblasts (EVTs). The number of granulysin-positive CD56(bright) NK cells increased significantly in the decidua basalis during spontaneous abortion compared with normal pregnancy; however, granzyme B- and perforin-positive cells did not change. Interestingly, the expression of granulysin was also detected in the nuclei of EVTs in spontaneous abortion samples. When IL-2-stimulated CD56(bright) NK cells were cocultured with EVT cells (HTR-8/SV40neo), granulysin was found initially in the cytoplasm and then accumulated in the nuclei of the HTR-8/SV40neo cells. Furthermore, transfected cells expressing a GFP-granulysin fusion protein induced apoptosis in HTR-8/SV40neo cells independently of caspases. Our results suggest that granulysin-positive uterine NK cells attack EVTs; subsequently, the uNK-derived granulysin actively accumulates in the nuclei of EVTs, causing the death of EVTs due to apoptosis. These data support a new apoptosis pathway for trophoblasts via uNK-derived granulysin, suggesting that granulysin is involved in spontaneous abortion.

Inadequate tolerance induction may induce pre-eclampsia.

The fetus is semi-allograft to the maternal host; therefore, a system of tolerance must be present during pregnancy. Epidemiological findings support a relationship between pre-eclampsia and the failure of tolerance induction. For induction of major histocompatibility complex (MHC) class I-specific tolerance, long-term exposure to seminal fluid, which contains paternal soluble MHC class I antigens, may induce paternal MHC class I-specific tolerance. Furthermore, soluble HLA-G1, which induces the deletion of CD8(+) T-cells, and the combination of maternal killer-immunoglobulin-like receptors (KIR) on NK cells and fetal HLA-C, which affects the balance between inhibition and activation signals of NK cells, regulatory CD8(+) T cells, and regulatory NK cells, may play very important roles in the induction of MHC class I-specific tolerance. On the other hand, exposure to sperm, which express paternal HLA-DR, and trophoblastic debris, which contain intracellular fetal HLA-DR, may induce paternal MHC class II-specific tolerance. In this process, CD4(+)CD25(+) regulatory T (Treg)-cells play central roles. In this review, we discuss the relationship between the risk of pre-eclampsia and tolerance induction.

The role of the immune system in preeclampsia.

Recent data demonstrate that an altered immune response may play a key role in the development of preeclampsia. Some epidemiological findings and animal models support this idea. In this article, we review the innate immune system and adaptive immune system in preeclampsia and discuss the pathophysiology of preeclampsia from an immunological viewpoint. The most characteristic immunological finding in preeclampsia is the activation of both the innate and adaptive immune system. Activated neutrophils, monocytes, and NK cells initiate inflammation which induce endothelial dysfunction, and activated T cells may support inadequate tolerance during pregnancy. The cytokine profile in preeclampsia shows that the production of type 1 cytokines, which induce inflammation, is dominant while the production of type 2 cytokines, which regulates inflammation, is suppressed. Furthermore, the immunoregulatory system is down-regulated in preeclampsia and persistent inflammation reduces regulatory T cell function. Therefore, systematical immunoactivation may be one cause of preeclampsia.

Interleukin-8 and glucose in amniotic fluid, fetal fibronectin in vaginal secretions and preterm labor index based on clinical variables are optimal predictive markers for preterm delivery in patients with intact membranes.

AIM: Various predictive markers for preterm delivery have been proposed in previous studies. We investigated which marker is most reliable. METHODS: In 126 patients with preterm labor before 32 weeks of gestation and intact membranes, who had regular uterine contractions with cervical changes effaced > or =50%, we evaluated seven markers: interleukin (IL)-8, glucose, and granulocyte count in amniotic fluid (AF); fetal fibronectin (fFN) in vaginal secretions; IL-8 in cervical mucus; cervical length; and preterm labor index (PLI) based on clinical variables. The relationships of these variables to the occurrence of preterm delivery before 34 weeks were examined by logistic regression analysis. RESULTS: Values for AF IL-8, AF granulocyte count, fFN in vaginal secretions, and PLI were significantly higher, while the value for AF glucose was significantly lower, in patients delivering before 34 weeks than those in patients delivering at or following 34 weeks (P < 0.0001 for all). The most sensitive marker for predicting delivery before 34 weeks was AF IL-8 (sensitivity, 67.8%). The most specific markers were AF IL-8 (specificity, 95.5%) and PLI (specificity, 95.5%). By the logistic regression analysis, AF IL-8, AF glucose, fFN in vaginal secretions and PLI showed independent relationships with delivery before 34 weeks (P = 0.0009, P = 0.0032, P = 0.0131 and P = 0.0038, respectively). CONCLUSIONS: In preterm labor with intact membranes, AF IL-8, AF glucose, fFN in vaginal secretions and PLI were highly predictive markers for detecting preterm delivery before 34 weeks.

Plasma granulysin concentrations and preeclampsia risk.

OBJECTIVE: Epidemiological, clinical and histological data suggest intriguing similarities between preeclampsia and graft-host-rejection. Granulysin, a novel biomarker of overall cellular immunity, is secreted by natural killer cells and cytotoxic T lymphocytes, which are associated with graft-host-rejection. Plasma granulysin was elevated in Japanese preeclamptic women. DESIGN AND METHODS: 50 preeclampsia cases and 50 normotensive controls (USA) were studied. Plasma granulysin at delivery was determined using enzyme immunoassay. Logistic regression procedures were used to estimate odds ratios (OR) and 95% confidence intervals (CI). RESULTS: Granulysin were elevated in preeclampsia cases compared with controls (3.01+/-0.18 vs. 2.22+/-0.14 ng/mL, p<0.01). After adjusting for age, body-mass-index and race, women with higher granulysin concentrations (> or =1.89 ng/mL) experienced a 2.9-fold (95%CI 1.1-7.8) increased preeclampsia risk compared with women with lower granulysin (<1.89 ng/mL). CONCLUSIONS: These data offer further evidence of a predominant Th1 immune status associated with preeclampsia. Prospective studies are needed to determine whether granulysin is elevated early in pregnancy.

Evaluation of effectiveness of prophylactic cerclage of a short cervix according to interleukin-8 in cervical mucus.

OBJECTIVE: This study was undertaken to compare rates of preterm delivery according to cervical mucus interleukin-8 (IL-8) among women who underwent cerclage because of a short cervix. STUDY DESIGN: This retrospective study included 16,508 patients whose cervical length and cervical mucus IL-8 concentrations were measured between 20 and 24 weeks. A short cervix was defined by a length of 25 mm or less, whereas IL-8 concentrations exceeding 360 ng/mL were considered high. Whether to perform cerclage was decided by clinicians without consideration of IL-8 concentrations. RESULTS: Among all subjects, a significantly smaller percentage of subjects avoided delivery before 37 weeks when cervical mucus IL-8 was elevated (P = .0302) or the cervix was short (P < .0001). Among patients with a short cervix, preterm delivery was more likely when cervical mucus IL-8 was elevated. Overall, risk of preterm delivery in patients with a short cervix did not differ between those undergoing and not undergoing cerclage. However, among patients with a short cervix, those with normal IL-8 concentrations in cervical mucus were less likely to have preterm delivery if they underwent cerclage (before 37 weeks, 33% vs 54.5%, P = .01; before 34 weeks, 4% vs 13.6%, P = .03). In contrast, when cervical mucus IL-8 was high, delivery before 37 weeks was more likely with than without cerclage (78% vs 54.1%, P = .03). CONCLUSION: With normal cervical mucus IL-8, cerclage treatment for cervical shortening may reduce the rate of preterm delivery, but with elevated cervical mucus IL-8 cerclage may be harmful.

The incidence of pre-eclampsia among couples consisting of Japanese women and Caucasian men.

Recent data from Hiby (2004) have suggested that a combination of maternal killer immunoglobulin receptor (KIR) AA genotype and fetal HLA-C2 genotype increases the risk of pre-eclampsia. Different human populations have a reciprocal relationship between KIR AA frequency and HLA-C2 frequency. Japanese people have highest frequency of KIR-AA alleles and lowest frequency of HLA-C2 alleles. However, Caucasians have a moderate frequency of KIR-AA and HLA-C2 alleles. If this hypothesis is correct, the incidence of pre-eclampsia among couples consisting of Japanese women and Caucasian men should be higher than that among couples consisting of Japanese women and Japanese men. Therefore, we investigated the incidence of pre-eclampsia among 324 couples consisting of Japanese women and Caucasian men. The incidence of pre-eclampsia in these couples consisting of Japanese women and Caucasian men was similar to that in Japanese women and Japanese men. Our data do not support that of Hiby et al. [Hiby, S.E., Walker, J.J., O'Shaughnessy, K.M., Redman, C.W.G., Carrington, M., Trowsdale, I., Moffett, A., 2004. Combinations of maternal KIR and fetal HLA-C genes influence the risk of pre-eclampsia and reproductive success. J. Exp. Med. 200, 957-965], although we did not check the haplotypes for HLA-C and KIR.

Fundamental study of a newly developed medium on detection of Lactobacillus.

OBJECTIVE: To investigate the specificity of a newly developed Lactobacillus medium (LB medium) and its usefulness for the detection of Lactobacillus. METHOD: Chlorophenol red was added to Rogosa SL Broth to make the LB medium whose pH was adjusted to 5.90. After incubation of Lactobacillus in LB medium for 24 hours, the correlation between the number of bacteria and the medium pH was examined. The change of pH was also examined when other microbes coexisted with Lactobacillus. RESULTS: Six species of Lactobacillus were tested. The medium pH decreased to 4 approximately 5 after incubation of any of these species for 24 hours. Though there were some differences in the acid-producing ability among species, a significant negative correlation (r = -0.93 approximately -0.99) was found between the initial number of bacteria and the medium pH with all species tested. The color of the medium did not change after incubation of other bacteria that might also be found in the normal vagina. The performance of the medium was not affected by the presence of large numbers of other microbes. In L. gasseri JCM 1131, the test microbe for color sample, the number of microbes whose color changed (red-->orange-->yellow) were approximately consistent with that of microbes reported in bacterial vaginosis (BV). CONCLUSION: This method can be easily applied to detect Lactobacillus species as the number of Lactobacillus can be estimated based on the change in color of the medium for less than 24 hours after incubation.

Cytokine profile of natural killer cells in early human pregnancy.

PROBLEM: To examine whether the NK1/NK2/NK3/NKr1 paradigm can be adapted in natural killer (NK) cells. METHOD OF STUDY: Mononuclear cells were isolated from peripheral blood and/or decidua in healthy non-pregnant women (n = 17), early pregnant women (6-12 weeks of gestation, n = 17) and miscarriage cases (6-11 weeks of gestation, n = 10). We investigated the production of transforming growth factor (TGF)-beta, interleukin (IL)-4, IL-5, IL-10, IL-13, interferon (IFN)-gamma and tumor necrosis factor-alpha from peripheral blood- and decidual-CD56bright NK cells and -CD56dim NK cells by flow cytometry. RESULTS: In the peripheral blood of the non-pregnant subjects, the main populations of CD56bright NK cells and CD56dim NK cells were IFN-gamma-producing NK1 type cells. Populations of IL-10-producing NKr1 type cells in peripheral blood CD56bright NK cells and CD56dim NK cells in early pregnant women were significantly greater compared with those in non-pregnant women, and these cells population decreased in miscarriage cases. In the early pregnancy decidua, the main populations of CD56bright NK cells and CD56dim NK cells were TGF-beta-producing NK3 type cells, and NK1 type cells were rare. NK3 type cells in decidua were significantly decreased in miscarriage cases compared with those in normal pregnant subjects. IL-4-, IL-5- or IL-13-producing NK2 type cells were rare in peripheral blood and decidua. CONCLUSION: These data support the NK1/NK2/NK3/NKr1 hypothesis. NKr1 type cells in peripheral blood and NK3 type cells in decidua might play some important roles in the maintenance of pregnancy by regulation of maternal immune function.

Novel ELISA for the detection of raw and processed egg using extraction buffer containing a surfactant and a reducing agent.

Enzyme-linked immunosorbent assay (ELISA) has been considered extremely useful for the detection of markers of allergenic substances in food, because it is simple, offers a suitable sensitivity, and is useful in providing quantitative results. Allergenic protein present in processed food can be denatured or altered, hindering therefore their possibility to be extracted and detected. This paper reports the development of an ELISA method that can be used for the determination of allergenic proteins in buffer solutions containing SDS, a surfactant, and 2-mercaptoethanol, a reducing agent. Measurement by ELISA in solutions containing 1% SDS and 7% 2-mercaptoethanol has been made possible by using an antibody prepared through immunization with an antigen denatured with SDS and 2-mercaptoethanol. This ELISA technique can be used to measure proteins in food that have been denatured by various manufacturing processes. An example is egg white albumin, which is susceptible to heat denaturation and has been difficult to recover from food in the past. Its recovery was improved 10- to 100-fold by the new ELISA method as compared with previous methods. This means that allergenic substances in food can now be detected quantitatively. This method can be very useful in allergy prevention and control strategies.

CD4(+)CD25high regulatory T cells in human pregnancy.

In both rodent and human systems, there is an emerging consensus that immunoregulatory activity specific for donor alloantigens is enriched in the CD4(+)CD25+ T cell population. The absence of CD4(+)CD25+ regulatory T (Treg) cells induces severe immunodeficiency with autoimmune disease, dermatitis and fatal infections in humans and mice. CD4(+)CD25+ Treg cells play a critical role in peripheral tolerance, transplantation tolerance and maternal tolerance to the fetus. Although both human and mouse CD4(+)CD25+ Treg have potent regulatory properties, surface phenotypes of human CD4(+)CD25+ Treg cells are not exactly the same as those of mouse CD4(+)CD25+ Treg cells. Murine CD4(+)CD25+ T cells are homogenous and exhibit regulatory function. On the other hand, CD4(+)CD25high T cells are the only cells which exhibit regulatory function in humans. Humans CD4(+)CD25low cells have no ability for immunosuppression. CD4(+)CD25high T cells inhibit the immunostimulation of conventional T cells through cell-to-cell contact or immunosuppressive cytokines such as interleukin 10 and transforming growth factor-beta. As another mechanism of immunosuppression, CTLA-4 on CD4(+)CD25+ regulatory T cells up-regulate indoleamine 2,3-dioxygenase (IDO) expression in dendritic cells which play important roles for immunosuppression. Here, we review the differences between humans and mouse Treg cells and the role of CD4(+)CD25+Treg during pregnancy.

Low adjusted serum ionized calcium concentration shortly after birth predicts poor outcome in neonatal hypoxic-ischemic encephalopathy.

AIM: Hypoxic-ischemic reperfusion injury causes either necrosis or apoptosis, and the influx of ionized calcium into cells is the major cause of both types of cell death. The aim of this study was to investigate whether or not the serum ionized calcium concentration in neonates with hypoxic-ischemic encephalopathy (HIE) could be used to predict their outcome. METHODS: Serum samples were obtained shortly after birth from 20 HIE neonates who had not urinated or received treatment with calcium. Serum ionized calcium concentrations were adjusted for pH using a correction formula. Twelve neonates without any disease were selected as a control. The results were compared between nine HIE neonates who made a full recovery, 11 who died or had neurologic deficits, and 12 normal neonates. RESULTS: Considered together, the two HIE groups had lower serum ionized calcium concentrations (1.05 +/- 0.10 mmol/L) than the control group (1.22 +/- 0.07 mmol/L; P < 0.0001). Moreover, serum ionized calcium concentrations in the group with the poor outcome (0.99 +/- 0.07 mmol/L) were lower than those in the group that made a full recovery (1.13 +/- 0.06 mmol/L; P=0.0016). CONCLUSIONS: The serum ionized calcium concentrations shortly after birth were significantly lower in neonates with HIE who had a poor outcome. Low concentrations may reflect multiple organ damage, particularly involving the brain.

IDO expression on decidual and peripheral blood dendritic cells and monocytes/macrophages after treatment with CTLA-4 or interferon-gamma increase in normal pregnancy but decrease in spontaneous abortion.

Recent data demonstrated that CD4+CD25+ regulatory T (Treg) cells and an enzyme called indoleamine 2,3-dioxygenase (IDO) mediate maternal tolerance to the fetus. Interestingly, Treg cells express the CTLA-4 molecule on their surface, and B7 (CD80/86) ligation by CTLA-4 enhanced IDO activity of dendritic cells (DCs) and monocytes by the induction of interferon gamma (IFN-gamma) production. In this study, we studied the IDO expression on peripheral blood monocytes and decidual monocytes or DCs after treatment with CTLA-4/Fc fusion protein or IFN-gamma using flow cytometry. IDO expressions on both peripheral blood DC and decidual DC and monocytes were up-regulated during normal pregnancy. On the other hand, both IDO expression on DC and monocytes after IFN-gamma treatment or CTLA-4 treatment were decreased in spontaneous abortion cases. The expression of CD86 on peripheral blood and decidual monocytes and DC in spontaneous abortion cases was lower compared with those in normal pregnancy subjects. Also, IFN-gamma production by decidual and peripheral blood mononuclear cells after CTLA-4/Fc treatment in spontaneous abortion cases was significantly lower than those in normal pregnancy subjects. These data suggest that CTLA-4 on Treg cells up-regulates IDO expression on decidual and peripheral blood DC and monocytes by the induction of IFN-gamma production.

Relationship between cervical mucus interleukin-8 concentrations and vaginal bacteria in pregnancy.

PROBLEM: High interleukin (IL)-8 concentration in cervical mucus in the second trimester is a risk factor for premature birth. We investigated the relationship between vaginal pathogens and IL-8 in cervical mucus. METHOD OF STUDY: In 501 women with single pregnancy, vaginal secretions were cultured for bacteria and cervical mucus IL-8 concentrations were measured between 20 and 24 gestational weeks. RESULTS: Lactobacillus species were detected in 56.0% of 84 subjects with high IL-8 (> or =377 ng/mL), significantly less often than in 417 subjects with IL-8 below 377 ng/mL (84.7%; P < 0.0001). Anaerobic pathogens were detected in 83.3% of high IL-8 subjects, significantly more often than in normal IL-8 subjects (43.9%; P < 0.0001). By multivariate analysis, cervical IL-8 was significantly high only in subjects without Lactobacillus species; they showed a significantly higher prematurity rate than Lactobacillus-positive subjects. CONCLUSIONS: Absence of vaginal Lactobacilli was associated with increased cervical IL-8 and increased risk of premature delivery.

Elevated interleukin-8 in cervical mucus as an indicator for treatment to prevent premature birth and preterm, pre-labor rupture of membranes: a prospective study.

PROBLEM: We investigated whether cervical shortening and high interleukin (IL)-8 in cervical mucus were valuable indications for treatment to prevent premature birth and preterm, pre-labor rupture of membranes (pPROM). METHOD OF STUDY: Pregnant women were divided into group A, in which neither cervical IL-8 nor cervical length was measured in the middle trimester; and groups B and C, in which cervical length and cervical IL-8 were measured, and bed rest or cerclage was performed when cervical shortening was detected. In group B, vaginal washing with povidone iodine and insertion of chloramphenicol vaginal tablets were carried out in women with IL-8 elevations. RESULTS: In group B, duration of pregnancy was significantly prolonged compared with group A and C, and occurrence of pPROM was significantly lower. No significant differences were found in those rates between groups A and C. CONCLUSION: Successful treatment for women with IL-8 elevations in cervical mucus decreased rates of premature birth or pPROM.