The long-term effects of endometrioma surgery on ovarian reserve: a prospective case-control study.

The objective of this study was to evaluate the long-term effects of endometrioma excision on ovarian reserve. This study evaluated the long-term effects of endometrioma excision on ovarian reserve. A total of 63 women were enrolled in this prospective case-control study; 21 women had histories of endometrioma surgery (study group), 21 women had diagnoses of endometrioma, and 21 healthy age-matched women served as controls. Participants were recruited from the Department of Obstetrics and Gynecology, Inonu University Faculty of Medicine, between January 2007 and January 2016. The mean follow-up duration after endometrioma surgery was 30.4 ± 18.0 months for the study group. The mean follicle-stimulating hormone, luteinizing hormone and estradiol levels were similar among groups, but the anti-Müllerian hormone (AMH) level was significantly lower in the surgery group than in the control group (p < .001). The mean AMH level was 42% lower in the endometrioma surgery group than in the endometrioma group and 30% lower in the endometrioma group than in the control group (p = .080 and p = .160, respectively). Endometrioma has a detrimental effect on ovarian reserve, and decreased ovarian reserve compared with that in healthy fertile subjects without endometrioma is evident shortly after endometrioma excision. However, the endometrioma excision procedure does not significantly decrease the ovarian reserve in the long term.

Synthesis of Cyclodextrin-Based Multifunctional Biocompatible Hydrogels and Their Use in the Prevention of Intrauterine Adhesions (Asherman's Syndrome) after Surgical Injury.

Asherman's syndrome, which can occur during the regeneration of damaged uterine tissue after surgical interventions, is a significant health problem in women. This study aimed to acquire and characterize cyclodextrin-based hydrogels, which can be used to prevent Asherman's syndrome, and investigate their effectiveness with biomedical applications. A series of hydrogels were synthesized from the cross-linking of ß-cyclodextrin and different polyphenols with epoxy-functional PEG. Their chemical, physical, and biological properties were subsequently determined. The results demonstrated that the cyclodextrin-based hydrogels had a porous structure, high swelling ratio, good injectability, drug release ability, and antioxidant activity. Cell culture results illustrated that the hydrogels had no significant cytotoxicity toward L929 fibroblast cells. Considering all properties, the ß-CD-PEG-600-Ec hydrogel showed the most satisfactory properties rather than other ones. The potential of this hydrogel in preventing Asherman's syndrome was evaluated in a rat model. The results revealed that the ß-estradiol- and melatonin-loaded cyclodextrin-based multifunctional hydrogel group both structurally and mechanically showed an antiadhesion effect in the uterus and a therapeutic effect on the damage with the ß-estradiol and melatonin that it contains compared to the Asherman (ASH) group. This double drug-loaded hydrogel can be a promising candidate for preventing Asherman's syndrome due to its versatile properties.

Salusins, newly identified regulators of hemodynamics and mitogenesis, increase in polycystic ovarian syndrome.

In the current study, we aimed to investigate whether serum salusin α and ß levels are different in PCOS subjects. Fifty women with PCOS and 50 healthy, age- and body mass index matched controls were included to the prospective cross-sectional study. All subjects underwent venous blood drawing on the early follicular phase after an overnight fasting. Serum salusin α and ß levels were measured with EIA, and ELISA respectively. The relationships between serum salusin levels and anthropometric and metabolic parameters were also assessed. Plasma salusin α and ß levels were higher in women with PCOS compared to control group. Serum salusin α level correlated positively with salusin ß and fasting serum insulin levels. The serum salusin ß levels were correlated positively with HOMA-IR, TG, LDL-C, LH, FSH, and total testosterone levels. Our results indicate that salusins, newly identified regulators of hemodynamics and mitogenesis, are increased within the serum of women with PCOS.

Maternal and fetal adropin levels in gestational diabetes mellitus.

AIM: To evaluate maternal and cord blood serum adropin concentrations in pregnant women with gestational diabetes mellitus (GDM). STUDY DESIGN: Twenty pregnant women with GDM and 20 gestational age-matched healthy pregnant women participated in the study. Maternal serum and cord blood adropin levels were assessed using an enzyme immunosorbent assay, at the time of birth. The relation of maternal serum and cord blood adropin levels with metabolic parameters were also assessed. RESULTS: The mean maternal and cord serum adropin in the GDM group were significantly lower than those of the control women (P=0.01 and P<0.001, respectively). Maternal serum adropin levels did not correlate with either fetal serum adropin levels or maternal metabolic values. CONCLUSION: The data suggest that low adropin levels may contribute to the underlying pathogenesis of GDM.

Amniotic fluid urocortin-1 concentrations for the prediction of preterm delivery.

AIM: The aim of this study was to analyze whether urocortin-1 concentration in midtrimester amniotic fluid could serve as an indicative marker of preterm labor. MATERIAL AND METHODS: A retrospective cohort study was conducted. Urocortin-1 concentrations in midtrimester amniotic fluid were measured in 22 pregnant women with preterm deliveries and 45 women who delivered at term using enzyme-linked immunosorbent assay. RESULTS: The median amniotic fluid urocortin-1 concentration was significantly lower in the women with preterm birth (40.06 pg/mL; range, 13.77-67.58 pg/mL) than in the women who gave birth at term (49.56 pg/mL; range, 26.25-175.9 pg/mL; P = 0.022). The result of receiver-operator curve analysis indicates that an amniotic fluid urocortin-1 concentration ≤ 57.88 pg/mL had an area under the curve of 0.673 (95% confidence interval, 0.55-0.78; P = 0.01) with a sensitivity of 81.8%, specificity of 40.0%, positive predictive value of 40%, and a negative predictive value of 82% in identifying which of the patients subsequently delivered prematurely. CONCLUSIONS: These results suggest that low urocortin-1 concentration in midtrimester amniotic fluid could be used as an indicative marker of preterm birth.

Maternal and fetal serum orexin-A levels in gestational diabetes mellitus.

AIM: Evidence suggests that orexin regulates food consumption, glucose metabolism and insulin secretion. Orexin may have a role in the pathogenesis of type II diabetes mellitus, however its role in gestational diabetes mellitus is not known. We aimed to assess maternal serum and cord blood orexin-A (OXA) concentrations in pregnant women with gestational diabetes mellitus (GDM). MATERIAL AND METHODS: Thirty-five pregnant women with GDM and 35 gestational-age-matched healthy pregnant subjects participated in the study. Maternal serum and cord blood OXA levels were measured with enzyme immunoassay at the time of birth. The correlations between maternal serum and cord blood OXA levels, anthropometric and metabolic parameters were also assessed. RESULTS: The mean maternal and cord serum OXA (1.16±0.37 and 1.35±0.20ng/mL, respectively) in the GDM group were significantly different from those of the controls (1.58±0.59 and 1.25±0.21ng/mL, respectively). The mean maternal fasting-glucose-to-OXA ratio was significantly higher in the GDM group. In the GDM group, the mean maternal serum OXA levels were similar in the insulin (n=24) and diet (n=11) treated cases, respectively (1.13±0.36ng/mL and 1.21±0.41ng/mL). Maternal serum OXA levels positively correlated with fetal serum OXA and maternal glucose levels. OXA concentrations in maternal serum were negatively correlated with the fasting glucose, fasting insulin and homeostasis model assessment insulin resistance index. CONCLUSIONS: Maternal serum OXA levels decrease, and fetal serum OXA levels increase in women with GDM.

Effect of ovarian stimulation with recombinant follicle-stimulating hormone, gonadotropin-releasing hormone agonist and antagonists, on follicular fluid stem cell factor and serum urocortin 1 levels on the day of oocyte retrieval.

PURPOSE: To compare the serum and follicular fluid (FF) concentrations of stem cell factor (SCF) as well as the serum urocortin 1 (UCN1) concentration in gonadotropin-releasing hormone antagonist (GnRH-ant) and gonadotropin-releasing hormone agonist (GnRH-a) protocols for controlled ovarian hyperstimulation (COH) in IVF patients. METHODS: Follicular fluids and blood samples of 42 infertile women undergoing COH for IVF-embryo transfer with either GnRH agonist (n = 22) or GnRH antagonist (n = 20) protocols from 2010 to 2011 were collected during oocyte retrieval. SCF concentrations of serum and FF were assessed by sandwich enzyme immunoassay using ELISA Kit for SCF kid. Serum UCN1 concentration were measured using commercially available enzyme-linked immunosorbent assay. RESULTS: Concentrations of serum UCN1, serum and FF SCF were similar in the two groups. The serum SCF levels correlated strongly with the follicular SCF levels (r = 0.770, p < 0.001). The mean implantation rate, biochemical and clinical pregnancy rate and live birth rate per cycle were also similar in the groups. CONCLUSIONS: These observations suggest that there is no significant difference in follicular microenvironment in terms of SCF and UCN1 between agonist and antagonist protocols.

Germline cells in ovarian surface epithelium of mammalians: a promising notion.

It is a long held doctrine in reproductive biology that women are born with a finite number of oocytes and there is no oogenesis during the postnatal period. However, recent evidence challenges this by showing the presence of germ line stem cells in the human ovarian surface epithelium (OSE), which can serve as a source of germ cells, and differentiate into oocyte like structures. Postnatal renewal of oocytes may have enormous therapeutic potential especially in women facing the risk of premature ovarian failure idiopathically or iatrogenically after exposure to gonadotoxic chemotherapy and radiation for cancer therapy.This article reviews current knowledge on germ line stem cells in human OSE.

A comparative study on oxidative and antioxidative markers of serum and follicular fluid in GnRH agonist and antagonist cycles.

OBJECTIVE: To determine whether concentrations of oxidative stress markers of follicular fluid and serum are different in GnRH agonist protocol from GnRH antagonist protocol. MATERIAL AND METHOD: This was a cross-sectional study. Eighty-four women undergoing controlled ovarian stimulation with either GnRH agonist (n = 39) or GnRH antagonist protocols (n = 45) for IVF/ICSI treatment were assigned by a physician. Blood was obtained at the time of oocyte retrieval, and follicular fluid (FF) from the mature follicles of each ovary was centrifuged and frozen until analysis. Malondialdehyde (MDA), nitric oxide (NO), protein carbonyl (PC), hydroxyl proline (OH-P), sodium oxide dismutase (SOD), reduced glutathione (GSH), glutathione peroxidase (GSH-Px), adenosine deaminase (ADA) and xanthine oxidase (XO) were assessed in the serum and follicular fluid of each participants. RESULTS: The mean serum concentrations of GSH-Px, GSH and MDA were lower in the GnRH antagonist group compared to GnRH agonist group, but mean serum SOD was higher in the GnRH antagonist group. The mean follicular SOD, ADA and NO were higher in GnRH antagonist group than GnRH agonist group. The IVF/ICSI outcomes were similar in both groups. CONCLUSION(S): GnRH antagonist protocol is associated with increased oxidative stress. The relation of GnRH analogues with oxidative stress and its implication in follicular growth needs to be addressed in further studies.

c-Kit proto-oncogene expression in endometrial hyperplasia and endometrial cancer.

PURPOSE: To evaluate the expression of c-kit (CD117) in endometrial hyperplasia and endometrial cancer. METHODS: Expression of c-kit in 10 normal endometrium, 18 simple endometrial hyperplasia, 16 complex endometrial hyperplasia (10 cases with atypia and 6 cases without atypia), and 6 endometrial cancer were investigated by immunohistochemistry. RESULTS: c-Kit expression decreased as the lesion progressed to endometrial cancer. Immunostaining was mostly focal and weak in the normal endometrium and was mostly diffuse and strong in the simple and complex endometrial hyperplasia. CONCLUSIONS: Simple and complex hyperplastic endometrial tissues express diffuse cytoplasmic staining for c-kit and the expression decreases with the progression of the lesion.

Measuring the status of maternal serum thiol/disulfide couples in the diagnosis and/or the determination of the severity of late-onset preeclampsia.

OBJECTIVE: Preeclampsia (PrE) is a pregnancy-related disorder. PrE affects the health of the mother and/or the fetus binomial with short and/or long-term consequences. The role of oxidant/antioxidant molecules and aberrant maternal inflammation in PrE has been documented. However, the importance of antioxidant molecules such as thiols has been poorly documented. In this research, a possible link between serum thiols levels and the diagnosis/severity of late-onset PrE (L-PrE) was investigated. MATERIALS AND METHODS: We examined maternal serum native thiols, disulfide, total thiols levels, and their ratios in pregnant women with (n = 51) and without L-PrE (n = 50). The levels of these three markers were measured using spectrophotometric assays and compared. RESULTS: There were significant differences in terms of serum native and total thiols levels between patients with L-PrE and healthy pregnant women (p = .001, p = .008, respectively). Disulfide levels were not different in either group (p = 0.729). There was no difference between total thiols, native thiols, disulfide concentrations, and their ratios in patients with mild (23 patients) and severe (27 patients) preeclampsia in L-PrE (p ≥ .05). A significant discriminative role of native and total thiols for the presence of L-PrE, with cutoff values of 175.86 µmol/L and 296.73 µmol/L, respectively, were revealed in ROC curve analysis. CONCLUSIONS: Lower concentrations of total/native thiols were linked with the development of L-PrE. However, there is still a need for more clinically useful biomarkers/molecules and management strategies in PrE.

Independent predictors of cardiovascular risk in polycystic ovarian syndrome.

We aimed to determine the independent predictors of cardiovascular risk in polycystic ovarian syndrome (PCOS). Ninety-one PCOS and 51 control patients were enrolled to our prospective cross sectional case-control study. In early follicular phase hormonal and lipid profile, fasting insulin and CRP (hs-CRP) levels and glucose levels on fasting and 2 h after the 75 g glucose intake were determined. Insulin resistance (IR) was evaluated with homeostasis model assessment and free testosterone was determined with free androgen index. PCOS was found to be associated with dyslipidemia, hyperandrogenism, IR and sub-clinical inflammation. The prevalence of overweight-obesity (41.8% vs. 25.5%, p = 0.038), IR (42.9% vs. 23.5%, p = 0.035) and glucose intolerance (15.38% vs. 1.96%, p = 0.043) were significantly higher in PCOS compared to control group. Independent predictors of the risk of elevated hs-CRP level were PCOS status (OR = 5, 95% CI: 1.55-16.14, p = 0.007) and high BMI (OR = 4.2, 95% CI: 1.2-14.2, p = 0.022) and high BMI (OR = 1.2, 95% CI: 1.05-1.4, p = 0.007) and of TC/HDL ratio was high BMI (OR = 1.21, 95% CI: 1.05-1.4, p = 0.009) and increasing age (OR = 1.11, 95% CI: 1.01-1.2, p = 0.04). The presence of PCOS, independent from obesity and IR, is the strongest predictor of elevated hs-CRP level. Obesity and advanced age further increases the cardiovascular risk in PCOS.

Evaluation of asymmetric dimethylarginine, nitric oxide levels and associated independent variables in obese and lean patients with polycystic ovarian syndrome.

OBJECTIVE: To evaluate the asymmetric dimethylarginine (ADMA) and nitric oxide (NO) levels in obese and lean patients with polycystic ovarian syndrome (PCOS) and find out their relation with hormonal and metabolic parameters. METHODS: Twenty-two obese, 18 lean patients with PCOS and 11 obese, 24 lean healthy control patients were enrolled prospectively. Plasma ADMA and NO levels and arginine/ADMA ratio were evaluated on 3rd day of menstrual cycle after at least 10 h overnight fasting. RESULTS: Plasma ADMA, NO levels and arginine/ADMA ratio were similar in the groups. ADMA level did not correlate with the hormonal and metabolic parameters in patients with PCOS. However, NO correlated inversely with fasting insulin (r =  -0.353, p = 0.041) and homeostasis model of insulin resistance (HOMA-IR) (r =  -0.379, p = 0.027). Arginine/ADMA ratio also correlated inversely with fasting insulin (r =  -0.339, p = 0.050). In multinomial regression analysis the risk of low NO was associated independently with high fasting insulin (OR = 1.19, 95% CI 1.001-1.42, p = 0.049) and high HOMA-IR in patients with PCOS (OR = 2.26, 95% CI 1.03-4.98, p = 0.042). CONCLUSIONS: Insulin resistance may be the underlying mechanism of endothelial dysfunction through NO pathway in PCOS.

GnRH agonist trigger for women with breast cancer undergoing fertility preservation by aromatase inhibitor/FSH stimulation.

Aromatase inhibitors can be utilized to minimize oestrogen exposure in breast cancer patients undergoing gonadotrophin stimulation. This retrospective-prospective study determined whether using a gonadotrophin-releasing hormone agonist (GnRHa) trigger instead of human chorionic gonadotrophin (HCG) would reduce oestrogen exposure and improve cycle outcomes in aromatase inhibitor cycles. Seventy-four breast cancer patients who desired fertility preservation, with normal ovarian reserve and < 45 years of age received letrozole 5mg/day plus recombinant FSH 150-300 IU/day for ovarian stimulation. Subjects either received HCG 5000-10,000 IU (n=47) or leuprolide acetate 1mg (GnRHa, n=27) as trigger. Oestradiol measurements were repeated 4 days after the trigger and subjects were evaluated for ovarian hyperstimulation syndrome (OHSS). In the GnRHa group, oestradiol concentrations dropped significantly after the trigger than the HCG group (P=0.013) and there was a lower incidence of OHSS. GnRHa trigger resulted in a higher number and percentage of mature oocytes and a higher number of cryopreserved embryos or oocytes compared with HCG. GnRHa trigger improves outcomes by increasing the yield of mature oocytes and embryos in aromatase inhibitor cycles and also decreases the post-trigger oestradiol exposure as well as OHSS risks in women with breast cancer.

Vitreous vascular endothelial growth factor level during gestation.

PURPOSE: To evaluate the vitreous VEGF level alterations and its correlation with its plasma level if any, in gestation. METHODS: The blood and vitreous sampling procedures were performed before, during (20-23 days of gestation), and after gestation (2 months after birth) from seven White New Zealand rabbits. Blood samples were centrifuged then supernatants and vitreous samples were stored at -80 degrees C until assay. Measurements of serum and vitreous VEGF(165) were done by ELISA. RESULTS: The median plasma and vitreous VEGF(165) concentrations were 36.61 pg/ml (range 19.17-40.30), 14.92 pg/ml (range 8.95-15.20); 58.30 pg/ml (range 32.60-11.53), 20.51 pg/ml (range 11.94-21.26); and 35.30 pg/ml (range 27.30-39.60), 13.05 pg/ml (range 9.33-16.04) before, during and after gestation, respectively. Both the plasma and the vitreous VEGF(165) concentration during gestational period were significantly higher than pre-gestational period (P = 0.028, P = 0.018) and post-gestational period (P = 0.043, P = 0.028). The intravitreal VEGF(165 )concentrations appeared to correlate with plasma VEGF(165) levels (r = 0.940, P = 0.002) only in the gestational period. CONCLUSION: Vitreous VEGF levels increase in positive correlation with plasma VEGF during pregnancy. Further prospective studies are needed to support this finding.

Short-term effect of tibolone on C-reactive protein in hypertensive postmenopausal women.

OBJECTIVE: To evaluate the effects of tibolone on the serum C-reactive protein (CRP) in hypertensive postmenopausal women. METHODS: We enrolled 45 postmenopausal patients with hypertension and 17 normotensive postmenopausal women. Inclusion criteria were surgical menopause, the presence of vasomotor symptoms, and normal mammogram within 1 year, the absence of documented coronary artery disease, and normal electrocardiography. Forty hypertensive women and 17 normotensive women completed the 3-month period. Twenty-one hypertensive women received tibolone, whereas 19 served as control. At baseline and at 3 months, blood lipids and CRP were evaluated. RESULTS: Changes in lipid profile and CRP in the hypertensive and normotensive control groups during 3 months were not statistically significant. Total cholesterol levels decreased significantly after 3 months of tibolone treatment. A significant increase in CRP values was observed in the tibolone group (p=0.001). CONCLUSION: This trial demonstrated that tibolone treatment induced a significant increase in CRP and a significant decrease in total cholesterol in postmenopausal hypertensive women.

Metabolic and carbohydrate characteristics of different phenotypes of polycystic ovary syndrome.

OBJECTIVE: To compare the prevalence of various metabolic and cardiovascular risk factors and insulin resistance between polycystic ovary syndrome (PCOS) patients with or without hyperandrogenism. MATERIAL AND METHODS: This is a retrospective cross-sectional study involving women with PCOS as diagnosed according to the Androgen Excess (AE) Society definition (n=504) and women with normoandrogenemic PCOS (n=183). Anthropometrics, lipid profile, glucose, insulin, oral glucose tolerance test (OGTT), and reproductive hormone levels were evaluated. RESULTS: Women with PCOS diagnosed according to the AE Society had a significantly higher prevalence of metabolic syndrome compared with the normoandrogenemic PCOS phenotype: odds ratio (OR) 2.95 [95% confidence interval (CI) 1.21-7.21]. There was no significant difference in the prevalence glucose intolerance test between the groups [OR: 2.15, 95% CI 0.71-6.56]. The prevalence of low high density lipoprotein (HDL)-cholesterol in the group under the AE-PCOS Society criteria was higher than that of the normoandrogenemic PCOS group [OR: 2.82, 95%CI 1.29-3.36]. CONCLUSION: The risks of metabolic syndrome and cardiovascular disease may vary among the phenotypes of PCOS based on the Rotterdam criteria. This new data may be of reference in informing women with PCOS, although further prospective studies are needed to validate this proposition.

Association of folic acid receptor α in maternal serum with neural tube defects.

OBJECTIVE: To evaluate whether serum folic receptor α levels are changed in women whose previous pregnancies were complicated with neural tube defects (NTDs). METHODS: This was a case-control study that included 41 women as the control group who had previously had at least one healthy pregnancy and 37 women as the study group who had a previous pregnancy complicated with NTDs. Blood samples were obtained from all of the participants six weeks after the termination of pregnancy or delivery of a baby. Serum folate receptor α concentrations were analyzed using a commercially available enzyme-linked immunosorbent assay (ELISA) kit. RESULTS: The mean concentrations of serum folate receptor α were significantly lower in the NTD cases compared to those in the control group (p = 0.02). There was no significant difference in mean serum folate titers between the NTD cases and the control group (p = 0.07). CONCLUSION: Low serum folic acid receptor α levels in the current study did not appear to be a regulatory marker of maternal folate homeostasis per se but rather a factor that contributed to the development of NTDs.

Maternal, fetal and perinatal characteristics of preeclampsia cases with and without abnormalities in uterine artery Doppler indexes.

OBJECTIVE: To compare the maternal and fetal characteristics and perinatal outcome in mild and severe preeclampsia cases with and without uterine artery Doppler abnormalities. METHODS: Two hundred and fifty-nine mild and severe preeclampsia cases were evaluated retrospectively. Doppler measurements were done in the section where uterine artery raised from the hypogastric artery. Pulsatility index above the 95th percentile of the corresponding gestational age was accepted as abnormal. RESULTS: In mild and severe preeclampsia cases with abnormal Doppler (AD), the rate of intrauterine growth restriction, preterm birth and low birth weight was higher than, but the neonatal intensive care unit stay was similar to the cases with normal Doppler. Base excess was higher in the AD group, in mild and severe preeclampsia. The rate of low Apgar score at 5 min and perinatal mortality was higher in the AD group, in the mild preeclampsia. The strongest independent predictor of the perinatal morbidity and mortality was the presence of prematurity and of the prematurity was the presence of abnormal uterine artery Doppler. CONCLUSIONS: Maternal and perinatal morbidity and perinatal mortality increase in mild to severe preeclampsia cases with abnormal uterine artery Doppler. The abnormal uterine artery Doppler increases the morbidity and mortality by increasing the risk of prematurity.

Assessment of early decline in the percentage of ß-hCG values between days 0 and 4 after methotrexate therapy in ectopic pregnancy for the prediction of treatment success.

OBJECTIVE: To evaluate percentage changes in serum beta-human chorionic gonadotropin (ß-hCG) values between days 0-1, 0-4 and 0-7 as an indicator of methotrexate therapy success in extra-uterine pregnancies. MATERIAL AND METHODS: Women with ectopic pregnancy treated with single-dose methotrexate therapy between January 2011 and August 2012 were assessed. Recorded data were reviewed electronically from patient files. All women (n=93) with an ectopic pregnancy treated medically with intramuscular methotrexate (50 mg/m(2)) were included. The percentage changes in serum ß-hCG levels from day 0 to 1, day 0 to 4 and day 0 to 7 were calculated for each case. RESULTS: The median ß-hCG values decreased between days 0 and 4 by 55.8%, and 89.6% of these cases were treatment successes. The median initial serum ß-hCG values were lower in women with successful treatment, but this was not statistically significant (p=0.11). CONCLUSION: A decline in serum ß-hCG values between days 0 and 4 appears to be the best predictor. It would be beneficial to determine whether a woman with an ectopic pregnancy treated with single-dose methotrexate administration will be treated successfully.