Association of maternal serum Netrin-1 and Netrin-4 levels with placenta accreta spectrum.

OBJECTIVE: This study aimed to evaluate maternal serum Netrin-1 and Netrin-4 levels in pregnancies complicated with placenta accreta spectrum. PATIENTS AND METHODS: This cross-sectional study enrolled 49 pregnant women with the diagnosis of placenta accreta spectrum as the study group. Gestational age-matched 30 uncomplicated pregnant women with prior cesarean delivery and normal placentation were randomly selected as the control group. Maternal serum Netrin-1 and Netrin-4 levels were measured between weeks 34 and 36 of gestation. RESULTS: There was no significant difference between the groups in terms of demographic characteristics. Maternal serum Netrin-1 levels were significantly lower in placenta accreta spectrum cases compared with those in the control group (p=0.038). There was no significant difference between the groups in terms of maternal serum Netrin-4 levels (p>0.05). There was a significant negative correlation between maternal serum Netrin-1 levels and the number of prior cesarean deliveries (r=-0.313, p=0.005). CONCLUSIONS: The observed decrease in maternal serum Netrin-1 levels in placenta accreta spectrum cases associated with increased angiogenesis might be one of the factors involved in the pathophysiology of this disease.

Association among lipopolysaccharide, the transforming growth factor-beta superfamily, follicular growth, and transcription factors in spontaneous bovine ovarian cysts.

The aim of this study was to investigate some of the growth and transcriptional factors originating from oocytes and granulosa cells in follicular fluid and to identify the relationships between the basic blood metabolite-metabolic hormones and intrafollicular lipopolysaccharide (LPS) concentrations. Thirty cows included in the study were allocated into 2 groups comprising 15 cows with healthy preovulatory follicles (cyclic cows) and 15 cows with confirmed cystic follicles. The ovaries and uteri of all cows were assessed by transrectal ultrasonographic examination. Blood serum samples were collected at 15, 25, 35, 45, and 55 d after calving for analysis of nonesterified fatty acids, ß-hydroxybutyrate, insulin, glucose, IGF-I, ACTH, and cortisol. Ovaries and uteri were examined using transrectal ultrasound. Vaginal discharge was evaluated on the same days. Follicular fluid was also aspirated on days 35-55 from the healthy preovulatory follicles and cystic follicles using a transvaginal ovum pickup method. The densitometric levels of inhibin-α, growth and differentiation factor (GDF-9), bone morphogenetic protein (BMP-6), and GATA-4 and GATA-6 proteins were analyzed by the Western blotting technique; the concentrations of antimullerian hormone (AMH), IGF-I, estradiol-17 beta (E2), and progesterone (P4) were determined by ELISA; and the concentrations of LPS in the follicular fluid were measured by the Limulus amebocyte lysate test. The serum insulin, ACTH, and cortisol concentrations were higher in cystic cows than cyclic cows, but serum IGF-I concentrations were lower in cystic cows. The IGF-I concentrations of cystic follicular fluids were lower, whereas AMH levels were significantly greater than those of healthy preovulatory follicular fluids. The cystic follicles had significantly lower expression levels of GDF-9, BMP-6, GATA-4, and GATA-6; in contrast, inhibin-α expression and LPS concentrations were significantly higher than in healthy preovulatory follicles. The proportion of pathologic vaginal discharge within 25 d postpartum in cystic cows were higher than in the cyclic group. In conclusion, it is suggested that intrafollicular dysregulation of the transforming growth factor-ß superfamily, growth, and transcriptional factors is affected by high intrafollicular LPS concentrations and systemic metabolic changes and these disturbances may be responsible for the generation of ovarian cysts.

Does serum anti-Müllerian hormone levels always discriminate presence of the ovaries in adult bitches? Comparison of two ELISA kits.

Anti-Müllerian hormone (AMH) is produced in the ovary, and thus, it is an excellent marker of follicle pool in females. Current interest is the clinical use of this parameter as a biomarker to assess presence or absence of an intact ovary and to diagnose ovarian remnant syndrome (ORS) following incomplete ovariohysterectomy (OHE) in bitches. The aim of this study was to evaluate serum AMH concentrations in bitches (n = 34) before and after OHE using two different commercial ELISA kits, one of which is based on detecting human AMH and the other is based on detecting human AMH and the other specified for canine AMH. Furthermore, serum AMH levels were also measured in six ORS cases to compare the diagnostic utility of the two different ELISA kits. Serum AMH concentrations measured using the human and canine kit prior to and after OHE were 0.32 ± 0.24, 0.006 ± 0.22 ng/ml (p < .001) and 12.08 ± 22.81, 9.55 ± 15.42 ng/ml (p = .868), respectively. Thus, the canine-based kit was not able to reveal the significant drop in serum AMH levels. In conclusion, the human-based ELISA kits successfully detected the drop in serum AMH concentrations. Reliable results can only be achieved from well-designed ELISA kits, and AMH levels might be a useful diagnostic tool for the evaluation of presence or absence of ovaries as well as for the detection of ORS cases in bitches.

Ultrasound-guided intrauterine insemination versus blind intrauterine insemination: a randomized controlled trial.

PURPOSE: This study was performed to determine the effects of ultrasound (US) guidance during intrauterine insemination (IUI) on pregnancy rate. MATERIALS AND METHODS: This study is a prospective randomized controlled trial which was performed in Women's Health Research and Education Hospital, Infertility Unit. The study enrolled 130 couples who were scheduled to undergo IUI. The couples were randomized according to a computer-generated list into two groups; 1) the ultrasound-guided IUI group included 64 couples (n = 64) treated for 99 cycles 2) blind IUI group included 66 couples (n = 66) treated for 104 cycles. All women underwent controlled ovarian stimulation before IUI. The study's main measurements were pregnancy rate per cycle; pregnancy rate per woman. RESULTS: The pregnancy rates were similar in both the ultrasound-guided (USG) (16.2%, 16/99) and non-ultrasound-guided (NUSG)(12.5%, 13/104) groups (p = 0.386). CONCLUSIONS: The present results suggest a routine ultrasound guidance during IUI is not essential as it does not increase pregnancy rates but it can be used in such cases to overwhelm some sort of difficulties.

Characterization of transforming growth factor beta superfamily, growth factors, transcriptional factors, and lipopolysaccharide in bovine cystic ovarian follicles.

The process of transformation of growing bovine follicles into cysts is still a mystery. Local expression of proteins or factors, including transforming growth factor ß, growth factors, and transcription factors, plays a central role in mammals. Therefore, in abattoir-derived cystic ovarian follicles and follicular fluid, the role of some transforming growth factor ß superfamily proteins, insulinlike growth factor-1 (IGF-1) and GATA-4 and GATA-6, were investigated. The relationship between intrafollicular lipopolysaccharide (LPS) and etiopathogenesis of ovarian cysts was also assessed. Data on the preovulatory follicle and the largest follicle (F1) were compared. The number of intrafollicular LPS-positive samples and LPS concentrations were higher in cysts. Immunohistochemical staining was mildly positive for IGF-1, inhibin alpha, and GATA-4 in thecal cells. Staining for anti-Müllerian hormone (AMH), growth differentiation factor-9, bone morphogenetic protein-6 (BMP-6), and GATA-6 was insufficient for their quantitation, and oocytes could not be stained for any of the proteins tested in the cystic follicles. Expression of BMP-6, inhibin alpha, and IGF-1 was moderately higher in granulosa cells of F1 follicles, and all the proteins were moderately expressed in granulosa cells in preovulatory follicles. However, loss of GATA-6 staining was significant in F1 follicles. Intrafollicular progesterone, IGF-1, and AMH concentrations in cysts and F1 follicles were significantly higher than those in preovulatory follicles. Western blot analyses revealed that follicular fluid inhibin-α was strongly expressed, whereas expression of growth differentiation factor-9, BMP-6, GATA-4 and GATA-6 was lower in cysts than in preovulatory follicles. Also, high intrafollicular AMH concentration and low BMP-6 expression were closely associated with cystic degeneration and atresia. In conclusion, immunohistochemical loss of BMP-6 and GATA-6 in the granulosa cells together with high intrafollicular LPS levels may play important roles in disruption of the ovulatory mechanism and steroidogenic reactions in type 2 cyst. Also, high intrafollicular AMH concentration along with low BMP-6 expression may be used as indicators of the bovine degenarative ovarian follicles.

The effects of EGF and IGF-1 on FSH-mediated in vitro maturation of domestic cat oocytes derived from follicular and luteal stages.

The objective of this study was to evaluate the influence of epidermal growth factor (EGF) and insulin like growth factor-I (IGF-1) on the in vitro maturation of cat oocytes recovered from follicular and luteal stage ovaries. Oocytes from follicular (n=580) and luteal (n=209) stages were harvested and divided into four groups, which were cultured in FSH-mediated maturation medium supplemented with: (1) EGF alone (25ng/mL); (2) IGF-1 alone (100ng/mL); (3) EGF+IGF-1 (25ng/mL EGF+100ng/mL IGF-I); or (4) no growth factor (control). The proportion of follicular stage oocytes reaching the metaphase II stage was significantly higher than that of oocytes obtained at the luteal stage in both control and study groups (p<0.001). The percentages of oocytes reaching the metaphase II stage during the follicular period were 62.6% in control; 70.9% in EGF; 72.8% in IGF-1, and 78.1% in EGF+IGF-1 groups, whereas the respective values for gametes collected from luteal stage ovaries were 12.5%, 17.5%, 12.5%, and 16.9%. Additionally, the differences between the study and control groups were significant in the case of follicular stage oocytes. Finally, supplementing the maturation medium with EGF and/or IGF-1 significantly enhanced the meiotic maturation of oocytes recovered from follicular stage ovaries. The present study also demonstrated that the combination of EGF and IGF-I provides an additional or synergic effect on meiotic maturation of oocytes recovered from the follicular stage.

First Report of Bacterial Blight of Pomegranate Caused by Xanthomonas axonopodis pv. punicae in Turkey.

Pomegranate (Punica granatum L.) is an increasingly important fruit crop that is widely cultivated in Turkey. Typical bacterial blight symptoms were observed since spring of 2011 in pomegranate orchards located in Antalya Province. Symptoms were characterized by dark brown, angular to irregularly shaped spots on leaves and fruit; cankers on stems, branches, and trunks; and split trunks. The pathogen was isolated from leaf spots on naturally infected plants showing typical symptoms onto yeast dextrose chalk agar. Bright yellow bacterial colonies were consistently isolated. Bacterial strains were characterized as gram negative, oxidase negative, catalase positive, tobacco hypersensitivity positive, and able to produce acid from L-arabinose, D-galactose, D-glucose, and D-mannitol but not from D-xylose. Pathogenicity of the representative bacterial strain Serik-4 was performed on 2-year-old pomegranate plants cv. Hicaz. Leaves were sprayed until runoff with bacterial cell suspensions containing 107 CFU/ml. Inoculated plants were covered with transparent plastic bags to maintain moisture for 48 h. Negative control plants were inoculated with sterile distilled water. Plants were then incubated in a greenhouse at 30°C for 14 days. Symptoms on leaves included dark brown, angular to irregularly shaped water soaked lesions along the veins of the inoculated plants 10 days after inoculation. No lesions developed on the control plants. The symptoms on inoculated plants were similar to those on naturally infected plants. Yellow bacterial colonies were re-isolated from the inoculated plants and identified as the same as the original strain by conventional tests and FAME analysis, thus fulfilling Koch's postulates. Fatty acid methyl ester profiling of the representative strain Serik-4 using GC-MIDI (Microbial Identification Inc, Newark, DE) identified the genus of the bacterium as Xanthomonas. The identity of Serik-4 was further confirmed by amplifying the 16S rRNA gene with the universal primers 27F and 1492R (3) and sequence analysis (GenBank Accession No. KM007073). The 16S rRNA gene sequences of Serik-4 was 99% identical to the corresponding gene sequences of the Xanthomonas axonopodis pv. punicae strain present in the NCBI database (JQ067629.1). High incidence of bacterial blight caused by X. axonopodis pv. punicae on pomegranate has been previously reported in India (2), Pakistan (1), and South Africa (4). To our knowledge, this is the first report of bacterial blight on pomegranate caused by X. axonopodis pv. punicae in Turkey. References: (1) M. A. Akhtar and M. H. R. Bhatti. Pakistan J. Agric. Res. 13:95, 1992. (2) R. Chand and R. Kishun. Indian Phytopathol. 44:370, 1991. (3) D. J. Lane. Page 115 in: Nucleic Acid Techniques in Bacterial Systematics, 1991. (4) Y. Petersen et al. Australas. Plant Pathol. 39:544, 2010.

Characteristics and prognosis of ovarian metastatic tumors: review of a single-institution experience.

BACKGROUND: To evaluate the clinico-pathological characteristics and role of surgery in patients with ovarian metastasis. MATERIALS AND METHODS: Clinical data from 51 patients with pathologically confirmed ovarian metastasis were reviewed. RESULTS: Ovarian metastasis accounted for 14% of all malignant ovarian neoplasms (51/364). Of the 51 metastatic ovarian tumor cases, 24 originated from gynecologic malignancies, while 27 originated from non-gynecologic malignancies. Optimal cytoreduction was performed in 88% and 37% of patients with metastases of gynecologic and non-gynecologic origin, respectively. Patients with ovarian metastasis had a two-year survival rate in 82% of the gynecologic group and 70% of the non-gynecologic group (p = 0.35). The five-year survival rate of patients with non-gynecologic tumor origin (29%) was significantly worse (p = 0.04) than the survival rates of those with tumors of gynecologic origin (61%). In the non-gynecologic group, the five-year survival rates were significantly different between patients who were performed optimal cytoreductive surgery vs those without this procedure (42% and 20%, respectively; p = 0.04). CONCLUSION: Although complete surgical resection is not achievable in approximately two-thirds of patients with metastases of non-gynecological origin, optimal tumor cytoreduction appears to improve survival, which is statistically significant in all patients with ovarian metastatic tumors.

Diagnosis and management of cesarean scar pregnancy.

OBJECTIVE: To evaluate the diagnosis and management modalities of cesarean scar pregnancy according to our experience. DESIGN AND SETTING: Retrospective study at the Women's Health Research and Education Hospital. PATIENTS: Six patients were diagnosed and treated for cesarean scar pregnancy (CSP) with dilatation and curettage, methotrexate (MTX), or laparatomy. RESULTS: One patient chose the surgical option due to her desire to have a tubal ligation. In the second case methotrexate was applied initially, but two weeks later suction curretage was applied due to abdominal pain and vaginal bleeding. Suction curettage was used as an initial treatment for four patients. There were not any complications in three of four patients. One patient had heavy vaginal bleeding which started after curettage. On ultrasonographic examination, increasing hemorrhage was seen between the uterus and the bladder so subtotal hysterectomy was performed. DISCUSSION: Ultrasound should be used effectivelly in evaluation of pregnant patients with previous cesarean deliveries. There is still no unique treatment modality for CSP, so treatment should be tailored for each patient. Before the 7th week, abortion should be considered. After the 7th week, MTX and/or surgical options should be preferred.

Molecular characterization of sour orange (Citrus aurantium) accessions and their relatives using SSR and SRAP markers.

Citrus production with its many varieties is of importance since it provides economically important products for Turkish exports. Sour orange is a rootstock commonly used for propagating the different scion varieties. Knowledge of the genetic diversity of the rootstock accessions would be useful in order to improve citrus breeding programs. We studied genetic relationships and diversity of 51 accessions of sour orange (Citrus aurantium) and their relatives using SSR (simple sequence repeat) and SRAP (sequence-related amplified polymorphism) molecular markers. Twenty-one SRAP primer combinations were tested on these accessions and relatives, producing 167 polymorphic fragments, with a mean of 8.0 and a mean polymorphism information content value of 0.47. Seventeen SSR primers also produced 30 polymorphic fragments, with a mean of 1.4 per primer and a mean polymorphism information content value of 0.39. The unweighted pair-group method with arithmetic average analysis using combined SSR and SRAP data showed a similarity range from 0.71 to 1.00 among the accessions. In the cluster analysis, sour orange relatives were indicated as a separate group from sour orange. 'Macrophylla' and 'Mexican lime' were the accessions most distinct (0.71) from the others. We conclude that genetic diversity in these sour orange accessions is lower and some of them were identical.

Peripartum hysterectomy in Turkey: a case-control study.

This was a retrospective review of all cases of peripartum obstetric hysterectomy performed at the Istanbul Bakirkoy Women and Children's Teaching Hospital in the period between January 2001 and September 2008. We included any women who required emergency hysterectomy to control major postpartum haemorrhage after delivery. During the study period, there were 91 cases of peripartum hysterectomy. Two controls per case were randomly selected from the remaining births by using the random table. The incidence of emergency peripartum hysterectomy was 0.67 in 1,000 deliveries. The main indication for emergency hysterectomy was uterine atony in 52 cases (57.1%). The most independent risk factors for emergency hysterectomy were multiparity (odds ratios (OR) 17.3, 95% confidence interval (95% CI) 8.7-34.6); caesarean delivery in index delivery (OR 6.7, 95% CI 3.8-11.9) and caesarean section for placental abruption (OR 3.8, 95% CI 0.4-33.4). Our study suggests that multiparity, primary or repeat caesarean deliveries for placental abruption are independently associated risks for peripartum hysterectomy and uterine atony is the still most common indication for peripartum hysterectomy in Turkey.