Polymorphism in intron 2 of the interleukin-1 receptor antagonist gene, local midtrimester cytokine response to vaginal flora, and subsequent preterm birth

OBJECTIVE: This study investigated the association between polymorphism in intron 2 of the interleukin-1 receptor antagonist gene, midtrimester vaginal microflora, vaginal interleukin receptor antagonist and interleukin-1beta levels and subsequent spontaneous preterm birth. STUDY DESIGN: Vaginal samples from 212 women, collected at 18-22 weeks' gestation, were analyzed for the polymorphism in intron 2 of the interleukin-1 receptor antagonist gene by polymerase chain reaction, qualitative and quantitative vaginal microflora, and interleukin-1beta and interleukin-1ra concentrations by enzyme-linked immunosorbent assay. Pregnancy outcome data were subsequently obtained. RESULTS: Carriage of intron 2 of the interleukin-1 receptor antagonist allele 2 (IL1RN * 2) was associated with an elevated vaginal pH in black ( P < .001) and white ( P = .005) women, a reduced interleukin-1beta response to anaerobic Gram-negative rods and/or Gardnerella vaginalis ( P < .01), and a decreased rate of spontaneous preterm deliveries (6% versus 18%, P = .02). In black women, IL1RN * 2 carriage was associated with increased anaerobic Gram-negative rods, Mycoplasma, and Peptostreptococci and decreased Lactobacilli colonization. CONCLUSION: IL1RN * 2 carriage was associated with a blunted proinflammatory interleukin-1beta response to abnormal vaginal flora. This property may decrease susceptibility to infection-related preterm birth.     

Changes in serum levels of heat shock protein 70 in preterm delivery and pre-eclampsia

AIM: The aim of this study was to investigate heat-shock protein (Hsp)70 as a novel marker to evaluate the curative effects of treatment for preterm delivery high-risk patients and pre-eclampsia. METHODS: After obtaining informed consent, serum samples were collected from 31 preterm delivery high-risk patients with a tocolysis index of three points or above (A), seven pre-eclampsia patients (P), 46 normal pregnant women (B), and seven non-pregnant women (C). Of the 31 preterm delivery high-risk patients, 15 had preterm delivery (Ap) and 16 had full-term delivery (Af). The levels of Hsp70 were measured using enzyme-linked immunosorbent assay. RESULTS: The Hsp70 levels in normal pregnant women were 8.6 +/- 1.9 ng/mL (first trimester), 5.5 +/- 1.0 ng/mL (second trimester) and 5.5 +/- 0.7 ng/mL (third trimester). There was no statistical difference in the Hsp70 levels between the three trimesters. The mean Hsp70 levels were 21.9 +/- 5.3 ng/mL (A), 35.3 +/- 9.6 ng/mL (Ap), 9.4 +/- 2.2 ng/mL (Af), 24.4 +/- 3.6 ng/mL (P), 6.1 +/- 0.6 ng/mL (B), and 2.4 +/- 0.6 ng/mL (C). Group Ap had significantly higher Hsp70 levels than group Af (P = 0.0112) and group B (P <0.0001). The duration of pregnancy after hospitalization for group Ap was significantly shorter than that for group Af (P=0.0088) and group B (P <0.0001). Group P also had significantly higher Hsp70 levels than group B (P <0.0001). CONCLUSION: Because Hsp70 levels were particularly high in treatment-resistant preterm delivery cases, Hsp70 may prove to be a useful marker for evaluating the curative effects of treatment for preterm delivery.     

Alteration of heat shock protein 70 expression levels in term and preterm delivery

Abstract

Objective: Heat shock protein 70 (Hsp70), a highly conserved cellular stress protein, is produced in every organism from bacteria to man. The purpose of this study was to determine the difference in Hsp70 concentrations between term and preterm deliveries.

Methods: In total, 30 healthy term delivery and 99 preterm delivery (PD) women were recruited, including 46 women with preterm labor and intact membranes (PTL) with or without an intra-amniotic infection (IAI) and 53 women with a preterm premature rupture of membranes (PPROMs) with or without IAI. The Hsp70 levels in the maternal and the umbilical cord sera were tested by an enzyme-linked immunosorbent assay (ELISA); the expression levels of Hsp70 in the placentas were determined by immunoblotting and real-time PCR.

Results: In contrast to the expression levels in normal term controls, Hsp70 expression levels were upregulated in PD; similar changes in Hsp70 expression levels were detected in PD maternal and umbilical sera. Patients with IAI were associated with a higher Hsp70 concentration than those without IAI.

Conclusion: These results suggest a probable contributing role of altered Hsp70 expression levels in the pathophysiology of PD.     

Decreased expression of the heat shock protein hsp70-2 is associated with the pathogenesis of male infertility

Objective: To examine the expression of the heat shock protein hsp70-2, and the possible relationship with the pathogenesis of male infertility.

Design: Prospective study.

Setting: Reproductive testing laboratory in a university hospital.

Patient(s): Men undergoing testicular biopsy during an investigation of subfertility.

Intervention(s): Testicular tissues were obtained from biopsies of men undergoing infertility evaluation and subdivided into three groups: normal testes, maturational arrest and Sertoli cell-only syndrome. Immunostaining and Western blotting techniques determined expression of the heat shock protein hsp70-2

Main Outcome Measure(s): Expression of the heat shock protein hsp70-2 in the testes.

Result(s): The experimental data demonstrated that the heat shock protein hsp70-2 was expressed in the normal and maturation arrest testicular specimens. The heat shock protein hsp70-2 was strongly present in the cytoplasm of spermatocytes and spermatides in the adluminal compartment of the seminiferous epithelium in normal testis. However, maturation arrest testis tissue demonstrated light staining in spermatocytes and spermatides, and Sertoli-only specimens demonstrated no staining for the heat shock protein hsp70-2. The Western blotting data showed a 70-kDa heat shock protein in the normal and maturation arrest testicular tissues, but not in the Sertoli-only tissues.

Conclusions: These results suggest that the heat shock protein hsp70-2 is expressed in spermatocytes and spermatides in normal and maturation arrest tissues. However, the expression of the heat shock protein hsp70-2 was low in maturation arrest, and no heat shock protein hsp70-2 was demonstrated in Sertoli-only specimens. Therefore the decreased expression of the heat shock protein hsp70-2 is associated with the pathogenesis of male infertility.     

Elevated serum 70 kDa heat shock protein level reflects tissue damage and disease severity in the syndrome of hemolysis, elevated liver enzymes, and low platelet count

OBJECTIVE: We have recently demonstrated that serum 70 kDa heat shock protein (Hsp70) levels are increased in the syndrome of hemolysis, elevated liver enzymes, and low platelet count (HELLP syndrome). The aim of the present study was to investigate in an independent, larger cohort of patients whether serum Hsp70 levels are related to laboratory markers of HELLP syndrome. STUDY DESIGN: The study population included 14 patients with HELLP syndrome. Serum Hsp70 levels were measured by enzyme-linked immunosorbent assay. The relationship between serum Hsp70 levels and laboratory markers of hemolysis, hepatocellular damage, renal insufficiency, inflammation or disseminated intravascular coagulation (DIC), as well as platelet count was investigated by calculating correlation coefficients, standardized regression coefficients and by principal component analysis. RESULTS: Serum Hsp70 levels showed a very strong correlation to the markers of hemolysis (plasma free hemoglobin level, serum lactate dehydrogenase activity, and total bilirubin level) and of hepatocellular injury (serum aminotransferase activities), supported also by principal component analysis. Furthermore, circulating Hsp70 concentration reflected the severity of HELLP syndrome as expressed by the significant inverse correlation to the lowest platelet count. By contrast, there was no relationship between serum Hsp70 levels and markers of inflammation, coagulation, fibrinolysis or renal insufficiency. CONCLUSION: Elevated serum 70 kDa heat shock protein level seems to reflect tissue damage (hemolysis and hepatocellular injury) and disease severity in patients with HELLP syndrome. However, further investigations are needed to determine the clinical relevance of these findings.     

A positive association between interleukin-1 receptor antagonist and insulin resistance in postmenopausal women

Objective: The purpose of this study was to determine whether higher circulating interleukin-1 receptor antagonist (IL-1Ra), an anti-inflammatory cytokine, was associated with insulin resistance in postmenopausal women.

Methods: We measured IL-1Ra concentrations in 160 naturally postmenopausal women without a history of diabetes mellitus. A Pearson coefficient was computed to assess the relationship between plasma IL-1Ra and homeostasis model assessment of insulin resistance (HOMA-IR). The association between HOMA-IR and IL-1Ra plasma level above the median was assessed by logistic regression. Linear regression was used to explore the determinants of IL-1Ra plasma levels.

Results: A significant positive correlation existed between IL-1Ra and HOMA-IR (r?=?0.42, p?p?=?.007).

Conclusions: Our results demonstrated a positive association between plasma IL-1Ra and insulin resistance in postmenopausal women. This analysis suggested that insulin resistance was an important determinant of circulating IL-1Ra for these women.     

细胞周期蛋白依赖性激酶2、热休克蛋白60在宫颈癌组织中的表达

目的 :研究人子宫颈癌组织中细胞周期蛋白依赖性激酶 (CDK) 2、热休克蛋白 (HSP) 60的表达状态 ,并探讨它们与宫颈癌的关系。方法 :应用SP免疫组织化学法研究 2 5例宫颈癌组织、11例宫颈癌旁组织及 16例正常宫颈组织中CDK2、HSP60的表达。结果 :CDK2、HSP60蛋白定位于宫颈癌细胞的胞质中 ,在宫颈癌、癌旁组织及正常宫颈组织中表达的阳性率分别为 76.0 %、2 7.3%、31.3%和 84 .0 %、4 5.5%、37.5%。CDK2及HSP60在宫颈癌组织中表达阳性率较癌旁组织及正常宫颈组织升高 ,差异有显著性 (P <0 .0 5)。结论 :CDK2在宫颈癌组织中的过度表达与宫颈癌的分化密切相关 ,可通过改变细胞周期促进细胞异常增生发挥作用 ,HSP60在宫颈癌组织中的过度表达与宫颈癌的分化密切相关。     

Association between subjective feelings of distress,plasma cortisol,anxiety, and depression in pregnant women

Objective

Psychosocial stressors are consistently associated with antenatal anxiety and depression, while the impact of cortisol has proved inconsistent. Most studies have focused either on psychological or physiological stress indices. We investigated both subjective and endocrinologic indices of distress in the same subjects.

Study design

We performed a cross-sectional study in late pregnancy in 79 women to investigate associations between the factors involved in anxiety and depression. Outcome measures were the Beck Depression Inventory (BDI), State-Trait Anxiety Inventory (STAI), Perceived Stress Scale, a Likert-like scale for fear of delivery, a structured interview to assess health and socioeconomic status, and fasting plasma cortisol levels measured in the morning.

Results

High BDI and STAI scores were associated with high levels of perceived stress and fear of delivery, but not with levels of plasma cortisol typical of the gestation phase. A multiple regression analysis revealed that subjective feelings of distress explained over 50% of the variation in BDI and STAI scores. Plasma cortisol was not a significant predictor of psychometric scores and did not show significant correlation with them in correlation analyses, and subjects with low and high cortisol levels showed similar psychometric scores.

Conclusion

Antenatal depression and anxiety were significantly associated with subjective feelings of distress but not with increased cortisol. This finding may be explained by the blunted cortisol stress responses characteristic of pregnancy. The mechanisms mediating the effects of subjective distress remain obscure: likely candidates include monoamine neurotransmission, and/or stress-induced changes in glucocorticoid receptor expression or distribution.     

Amniotic fluid heat shock protein 70 concentration in histologic chorioamnionitis,term and preterm parturition

Objective. Heat shock protein (HSP) 70, a conserved member of the stress protein family, is produced in almost all cell types in response to a wide range of stressful stimuli, and its production has a survival value. Evidence suggests that extracellular HSP70 is involved in the activation of the innate and adaptive immune response. Furthermore, increased mRNA expression of HSP70 has been observed in human fetal membranes following endotoxin stimulation. This study was conducted to determine the changes in amniotic fluid HSP70 concentrations during pregnancy, term and preterm parturition, intra-amniotic infection (IAI), and histologic chorioamnionitis.

Study design. A cross-sectional study was conducted in 376 pregnant women in the following groups: (1) women with a normal pregnancy who were classified into the following categories: (a) women in the mid-trimester (14–18 weeks) who underwent amniocentesis for genetic indications and delivered normal infants at term (n=72); (b) women at term not in labor (n = 23); and (c) those at term in labor (n = 48). (2) Women with spontaneous preterm labor and intact membranes who were subdivided into the following categories: (a) preterm labor who delivered at term without IAI (n = 42); (b) preterm labor who delivered preterm without IAI (n = 57); and (c) preterm labor and delivery with IAI (n = 30). (3) Women with preterm prelabor rupture of membranes (PROM) with (n = 50) and without (n = 54) IAI. Among patients with preterm labor with intact membranes and preterm PROM who delivered within 72 hours of amniocentesis, placenta, umbilical cord, and chorioamniotic membranes were collected and assessed for the presence or absence of acute inflammatory lesions in the extraplacental membranes (histologic chorioamnionitis) and/or umbilical cords (funisitis). HSP70 concentrations in amniotic fluid were determined using a sensitive and specific immunoassay. Non-parametric statistics were used for analysis. A p value of <0.05 was considered statistically significant.

Results. Immunoreactive HSP70 was detected in 88% (332/376) of amniotic fluid samples. The median amniotic fluid HSP70 concentration was significantly higher in women at term without labor than in those in the mid-trimester (term no labor: median 34.9 ng/mL, range 0–78.1 ng/mL vs. mid-trimester; median 6.6 ng/mL, range 0–20.8 ng/mL; p<0.001). Among patients with spontaneous preterm labor and preterm PROM, those with IAI had a significantly higher median amniotic fluid HSP70 concentration than those without IAI (preterm labor with IAI: median 82.9 ng/mL, range 0–500 ng/mL vs. preterm labor without IAI: median 41.7 ng/mL, range 0–244 ng/mL; p = 0.001; preterm PROM with IAI: median 86.5 ng/mL, range 0–428 ng/mL vs. preterm PROM without IAI: median 55.9 ng/mL, range 14.9–299.9 ng/mL; p = 0.007). There was no significant difference in the median amniotic fluid HSP70 concentration between patients with preterm labor who delivered preterm without IAI and those who delivered at term (p = 0.6). However, among patients with preterm labor without IAI, there was an inverse relationship between amniotic fluid concentration of HSP70 and the amniocentesis-to-spontaneous delivery interval (Spearman's Rho = ?0.26; p = 0.02). Patients with histologic chorioamnionitis/funisitis had a significantly higher median amniotic fluid HSP70 concentration than those without inflammation (inflammation: median 108.7 ng/mL, range 0–500 ng/mL vs. without inflammation: median 67.9 ng/mL, range 7.1–299.9 ng/mL; p = 0.02). Women at term in labor had a median amniotic fluid concentration of HSP70 significantly higher than those not in labor (term in labor: median 60.7 ng/mL, range 0–359.9 ng/mL vs. term not in labor: median 34.9 ng/mL, range 0–78.1 ng/mL; p = 0.02).

Conclusions. Intra-amniotic infection, histologic chorioamnionitis, and term parturition are associated with elevated amniotic fluid HSP70 concentrations. HSP70 plays a role in the host defense mechanism by activating the innate arm of the immune response in women with intrauterine infection. The mechanisms of preterm and term parturition in humans may involve extracellular HSP70.     

Detection of the human 70-kD and 60-kD heat shock proteins in the vagina: relation to microbial flora, vaginal pH, and method of contraception

The expression of the 60-kD and 70-kD heat shock proteins (hsp60 and hsp70) in the vaginas of 43 asymptomatic women of reproductive age with or without a history of recurrent vulvovaginitis (RVV) were compared. Vaginal wash samples were obtained and assayed by enzyme-linked immunosorbent assay (ELISA) for human hsp60 and hsp70. Heat shock protein 70 was not detected in any of the 19 women with no history of RVV, and hsp60 was present in only one woman in this group. In contrast, in the RVV group, 11 (45.8%) were hsp60-positive and eight (33.3%) were hsp70-positive. The presence of either heat shock protein in the vagina was associated with an elevated vaginal pH (>4.5). Bacterial vaginosis or Candida was identified in some of the asymptomatic subjects; their occurrence was significantly higher in women with vaginal hsp70 than in women with no heat shock proteins. Oral contraceptives were used by 35.7% of subjects who were negative for vaginal heat shock proteins, as opposed to only 12.5% of women who were positive for hsp70 and 8.3% who were positive for hsp60. Expression of heat shock proteins in the vagina may indicate an altered vaginal environment and a susceptibility to vulvovaginal symptoms.     

Polymorphisms in the tumor necrosis factor-alpha gene at position -308 and the inducible 70 kd heat shock protein gene at position +1267 in multifetal pregnancies and preterm premature rupture of fetal membranes

OBJECTIVE: The purpose of this study was to determine the relationship between preterm premature rupture of membranes, tumor necrosis factor-alpha, and heat shock protein-70 gene polymorphisms in multifetal gestations. STUDY DESIGN: Buccal swabs from 101 mother-neonate pairs of multifetal pregnancies were tested for single nucleotide polymorphisms at position -308 of the tumor necrosis factor-alpha gene and +1267 of the heat shock protein-70 gene. Pregnancy outcome data were obtained subsequently. RESULTS: Tumor necrosis factor-alpha allele 2 carriage by the first-born occurred in 10 of 27 pregnancies (37.0%) that resulted in preterm premature rupture of membranes compared with 6 of 67 pregnancies (9.0%) without preterm premature rupture of membranes ( P = .002). The allele frequency of tumor necrosis factor-alpha allele 2 and heat shock protein-70 allele 2 in the first born was higher in pregnancies that were complicated by preterm premature rupture of membranes (18.5% vs 4.5%; P = .003; and 57.7% vs 41.3%; P = .04, respectively). There was no relationship between tumor necrosis factor-alpha allele 2 or heat shock protein-70 allele 2 carriage by the second fetus or mother and preterm premature rupture of membranes. CONCLUSION: Tumor necrosis factor-alpha allele 2 and/or heat shock protein-70 allele 2 carriage by the first-born fetus is associated with preterm premature rupture of membranes in multifetal pregnancies.     

脑缺血预处理对新生鼠海马CA1区突触素及热休克蛋白70的影响

目的探讨脑缺血预处理对新生Wistar鼠脑CA1区突触素和热休克蛋白70(HSP70)变化的影响。方法通过阻断7日龄新生Wistar大鼠右侧颈总动脉制备脑缺血模型,设置假手术组、缺血再灌注组、预缺血-缺血再灌注组。用免疫组化法检测三组新生鼠不同时点海马CA1区脑组织神经细胞突触素和HSP70的动态变化,并光镜下观察脑组织病理改变。结果突触素:缺血再灌注组和预缺血-缺血再灌注组海马CA1区突触素表达在再灌注后第3天开始增加,分别为10.76±0.99、15.89±0.59,7 d时达高峰,分别为13.14±0.76、18.33±1.11,14 d时分别为12.01±0.71、15.13±1.10,仍高于假手术组(8.31±0.01),三组比较差异均有统计学意义(P<0.05);HSP70假手术组海马CA1区有极少量HSP70表达。缺血再灌注组和预缺血-缺血再灌注组海马CA1区HSP70表达在再灌注后6 h开始增高,分别为1.48±0.82、1.90±0.10,24 h达高峰,分别为8.15±0.99、11.32±0.96,3 d时开始下降,分别为3.42±0.21、5.31±0.51,三组比较差异均有统计学意义(P<0.05)。预缺血组海马CA1区病理改变较缺血再灌注组轻。结论脑缺血预处理对再次脑缺血所致脑神经细胞损伤有保护作用;新生鼠脑缺血后3~14 d海马CA1区神经细胞突触素表达增加可反映受损神经细胞突触再生,说明受损神经细胞具有代偿再生重塑功能;脑缺血6 h后HSP70表达增多可能与神经细胞内源性保护机制有关。     

Association between uncoupling protein 2, adiponectin and resting energy expenditure in obese women with normal and low resting energy expenditure

Obesity is recognized as the most prevalent metabolic disease worldwide. Decreases in energy expenditure may increase risk of obesity. One of the key regulators of energy balance is uncoupling protein2 (UCP2), a transporter protein presents in mitochondrial inner membrane. Moreover, adiponectin is the most abundant adipocytokine, it may play a role in energy metabolism and gene expression of UCP2. The aim of this study was to investigate potential associations between the level of uncoupling protein 2 and adiponectin and their relationship with REE (Resting Energy Expenditure) in obese women with normal and low resting energy expenditure. A total of 49 subjects (women, 25–50 years old), were included in current study, 16 subjects with BMI?>?30 and low resting energy expenditure, 17 subjects with BMI?>?30 and normal resting energy expenditure and 16 non-obese subjects as a control group. Anthropometric, body composition parameters and resting energy expenditure were measured. Plasma adiponectin, UCP2 protein and total protein in PBMC were determined. Measured resting energy expenditure in obese subjects with low REE was significantly lower than other groups. Plasma adiponectin in the obese subjects with low REE was significantly lower compared to normal weight group. There was a significant relationship between ‘UCP2 protein/Total protein’ ratio and plasma adiponectin in obese group with low REE and in three groups when we pooled. There was a significant association between REE and plasma adiponectin in three groups when we pooled. There was a significant association between plasma adiponectin and REE. Moreover, there was a significant relationship between UCP2 and REE.     

Interleukins-1, -4, -6, -10, tumor necrosis factor, transforming growth factor-beta, FAS, and mannose-binding protein C gene polymorphisms in Australian women: Risk of preterm birth

OBJECTIVE: The purpose of this study was to examine the relationship between preterm birth and 22 single nucleotide polymorphisms in genes that encode cytokines and mediators of apoptosis and host defense. STUDY DESIGN: Two hundred two white women with a spontaneous preterm birth of <35 weeks of gestation were compared with 185 white women with term births. Genotyping was performed with polymerase chain reaction and sequence specific primers. Multivariable analyses included demographic and genetic variables. RESULTS: Alcohol (multivariable odds ratio, 2.3; P = .001] and substance use (multivariable odds ratio, 3.7; P = .01) were associated with preterm birth at <35 weeks of gestation. Smoking (multivariable odds ratio, 2.3; P = .03), haplotypes IL10 -1082A/-819T/-592A (multivariable odds ratio, 2.1; P = .04), tumor necrosis factor ( TNF )+488A/-238G/-308G (multivariable odds ratio, 2.4; P = .04), and IL4 -509C/C (multivariable odds ratio, 3.4; P = .02), and the presence of MBL2 codon 54Asp (multivariable odds ratio, 2.3; P = .02) were associated independently with preterm birth at <29 weeks of gestation. Homozygosity for IL10 -1082G/-819C/-592C haplotype (multivariable odds ratio, 1.9; P = .02) was more common in women with preterm premature rupture of membranes. CONCLUSION: Polymorphisms in immunoregulatory genes may influence susceptibility to preterm birth or premature rupture of membranes.