The long pentraxin-3 is a useful marker for diagnosis of ovarian torsion: An experimental rat model.

Pentraxin-3 (PTX3) plays an important role in the primary inflammatory response. We aim to evaluate PTX3 as a diagnostic marker of ovarian torsion in an experimental rat model. In this study, 16 female Sprague Dawley albino mature rats were randomly allocated to Group 1 (control, sham operated) and Group 2 (experimental ovarian torsion model). A torsion model was set up using atraumatic vascular clips just above and below the right ovary for a 3-h ischaemia. Blood samples were collected before and three hours after ovarian torsion. Three hours after ovarian torsion, right ovary was surgically removed for histopathological examination in both groups. There was no significant difference in preoperative PTX3 level in both groups (1.05 ± 0.20 ng/mL vs 1.09 ± 0.28 ng/mL, p > 0.05). Three hours after the operation, mean plasma level of PTX3 was significantly higher in ovarian torsion group than the control group (2.13 ± 0.49 ng/mL vs 1.07 ± 0.22 ng/mL, p = 0.001). Also, the mean total histopathological score was significantly increased in the torsion group. PTX3 can be used in clinical practice as a useful marker for early diagnosis of ovarian torsion.

Clinical significance of ASCUS and ASC-H cytological abnormalities: a six-year experience at a single center.

BACKGROUND: To evaluate colposcopic biopsy results of patients with cervical cytological findings of atypical squamous cells of undetermined significance (ASC-US) and atypical squamous cells with high-grade lesions that cannot be excluded (ASC-H). MATERIALS AND METHODS: A retrospective evaluation of data from 358 patients, who had cervical cytological findings of ASC-US (n = 335) and ASC-H (n = 23), and had colposcopic assessments between 2005 and 2011. RESULTS: Cervical biopsy results of patients diagnosed with ASC-US cytology (n = 335) revealed cervical squamous cell carcinoma 0.9 % (n = 3) at biopsy, cervical intraepithelial neoplasia 3 (CIN 3) in 3.8 % (n = 13), cervical intraepithelial neoplasia 2 (CIN 2) in 1.1 % (n = 4), cervical intraepithelial neoplasia 1 (CIN 1) in 35.2% (n = 118), and benign lesions in 59 % (n = 197). Cervical biopsy results of patients diagnosed with ASC-H cytology (n = 23) revealed CIN 3 at biopsy in 39.3% (n = 9), CIN 2 in 21.7% (n = 5), CIN 1 in 26% (n = 6), carcinoma in situ in 8.7% (n = 2), and squamous cell cancer in one patient (4.3%). CONCLUSION: The cytological diagnosis of ASC-US may lead to the diagnosis of cervical intraepithelial lesion of higher grades as well as cervical cancer and should be evaluated by colposcopic cervical biopsy.

The effects of sunitinib on endometriosis.

The aim of the present study was to evaluate the effect of sunitinib on endometriotic implants and adhesions in a rat endometriosis model. An experimental endometriosis model was created in 21 rats. These rats were randomly divided into three groups: Group 1 (control group, 7 rats) was given no medication; Group 2 (sunitinib group, 7 rats) was given 3 mg/kg per day of oral sunitinib; and Group 3 (danazol group, 7 rats) was given 7.2 mg/kg per day of oral danazol. The volume of endometriotic implants was calculated. The extent and severity of adhesions were evaluated. The groups were compared by the Student's t-test, analysis of variance (ANOVA) and the Mann-Whitney U test. There was no statistically significant difference in the mean volume of endometriotic implants before medication between three groups. The volume of implants and extent, severity, total score of adhesions were significantly decreased after medication in Group 2 and Group 3. We noted that the volume of the endometriotic implants and adhesion formation were decreased both after sunitinib and danazol treatment. As a result, sunitinib seems to be effective for endometriotic peritoneal lesions. The effects of sunitinib in rat models give hope for improving the treatment of human endometriosis and prevention of pain symptoms.

The determination of high-risk pregnancy: the use of antenatal scoring system.

AIMS: A standardized antenatal scoring system is not available in order to detect high-risk pregnancies at primary health care units in Turkey. The authors aimed to evaluate the applicability of the "Knox scoring system" in Turkey. MATERIALS AND METHODS: One hundred and twenty-nine pregnant women were included in study. They were assessed upon admission and then the same women were reassessed at the onset of labor by Knox scoring system. RESULTS: The Knox scoring system identified 65 pregnant women (50.4%) as high-risk upon admission while 22 pregnant women (17.1%) as high-risk at the onset of labor. Twelve pregnant women (9.3%) (one case of perinatal death and 11 cases of perinatal morbidity) had poor perinatal outcomes during the study period. The Knox scoring form administered upon admission yielded 58% sensitivity, 50% specificity, and 10% positive predictive value, while 91% sensitivity, 90% specificity, and 50% positive predictive value at the onset of labor. The diagnostic value of the Knox scoring system for determining high-risk pregnancies was not found statistically significant regarding admission (ROC value: 0.655; p > 0.05), while statistically significant regarding the onset of labor (ROC value: 0.946; p < 0.05). CONCLUSIONS: The use of the Knox scoring system for determining high-risk pregnancies seems to be effective at the onset of labor.

Pegylated liposomal doxorubicin in heavily pretreated epithelial ovarian cancer patients.

PURPOSE: While most patients with ovarian cancer respond to first-line treatment, 50-75% of these patients will eventually relapse. Pegylated liposomal doxorubicin (PLD) is an active agent indicated for the treatment of patients with disease that is refractory to both paclitaxel- and platinum-based regimens, but skin toxicity remains the dose-limiting toxicity of the drug. The primary objective of this retrospective study was to evaluate the activity and safety of this agent in patients with heavily pretreated ovarian cancer. PATIENTS AND METHODS: Patients with platinum-refractory/ resistant, paclitaxel-pretreated epithelial ovarian carcinoma were treated with PLD 50 mg/m2 in 4-week courses until disease progression or unacceptable toxicity. All patients had progressive disease (PD) before starting PLD. Primary endpoints were response rate, progression free survival (PFS) and toxicity and secondary endpoints duration of response (DOS) and overall survival (OS). RESULTS: Seventeen heavily pretreated patients (median number of previous chemotherapy regimens 3, range 1-5) with taxane- and platinum-refractory disease were analysed. No complete response (CR) was achieved, while 3 (17%) partial responses (PR) and 2 (11%) cases with stable disease (SD) were observed. The median PFS was 15 weeks (range 10-21) and median OS 32 weeks (range 16-47). Palmar plantar erythrodysesthesia (PPE) occurred in 4 (23%) patients and was of grade 4 in 1 (6%) patient. Stomatitis occurred in 3 (17%) patients and was grade 3 in 1 (6%) patient. Grade 3-4 neutropenia occurred in only 2 (12%) patients. No febrile neutropenia was encountered. CONCLUSION: Pegylated liposomal doxorubicin is an active and tolerable agent in heavily pretreated epithelial ovarian cancer patients.

Immunohistochemical evaluation of cell proliferation and apoptosis markers in ovaries and uterus of tamoxifen-treated rats.

The aim of the study was to evaluate the immunohistochemical expression of cell proliferation and apoptosis markers in the ovaries and uterus of tamoxifen-treated rats. Twelve rats (150-200 g) were divided into two equal groups. The study group received daily intraperitoneal injections of tamoxifen dissolved in 5% dimethyl sulfoxide (n= 6). The control group received only the vehicle (n= 6). The rats were sacrificed at the 20th day of injection and were perfused. The ovaries and uterus of the rats were extracted. The sections were immunohistochemically stained with cell proliferation marker Ki-67 and the apoptosis markers PTEN and CD95. The expressions of the markers were quantified by a semiquantitative H-score method in myometrium, endometrial glands, ovarian surface epithelium, ovarian follicles, corpus luteum, and ovarian stroma separately. The mean H-scores of CD95 and PTEN obtained from myometrium, glandular endometrium, ovarian surface epithelium, ovarian follicles, corpus luteum, and ovarian stroma did not show significant difference between the study and the control groups. Proliferative index (Ki-67) of endometrial glands was significantly higher in the study group than in the control group (P < 0.05). In addition, proliferative index (Ki-67) of corpus luteum was significantly higher in the study group than in the control group (P < 0.05). Tamoxifen treatment has a potential to stimulate the cell proliferation of endometrial glands and corpus luteum in tamoxifen-treated rats. Apoptosis markers of PTEN and CD95 did not demonstrate significant difference after the tamoxifen treatment.

Arsenic trioxide-loaded, microemulsion-enhanced cytotoxicity on MDAH 2774 ovarian carcinoma cell line.

The antiproliferative effect of As(2)O(3)-loaded microemulsion (As(2)O(3)-M) on human MDAH 2774 ovarian cancer cells was compared with a regular solution of the As(2)O(3). We used MDAH 2774 as model cell lines for ovarian cancer. The (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide) (XTT) and trypane blue dye exclusion tests were used to evaluate cytotoxicity. Apoptotic effect of solutions was evaluated using cell death detection kit. Standard microemulsion formulation used in this experiment contains 5 x 10(-6) M As(2)O(3). It was clearly demonstrated that As(2)O(3)-M had a significant cytotoxic effect on MDAH 2774 cell line, and the cytotoxic effect of As(2)O(3)-M was significantly higher than that of regular As(2)O(3) solutions. Even approximately 6000 times diluted microemulsion formulation loaded with 5 x 10(-6) M As(2)O(3) showed a cytotoxic effect. As a result, this diluted concentration (approximately 8 x 10(-10) M) was found to be approximately 6000 times more effective than regular As(2)O(3) solutions (5 x 10(-6) M). Moreover, this diluted concentration resulted in 1.5-fold enhancement of apoptosis. According to the in vitro cytotoxicity studies, we concluded that by incorporating As(2)O(3) into the microemulsion (As(2)O(3)-M), which is a new drug carrier system, it is possible to increase antiproliferative effect of regular As(2)O(3) on MDAH 2774 cells. Translating these results to in vivo conditions would open new windows in the treatment of ovarian cancer.

How can we predict ureteral obstruction after gynecological surgery? The changes in Doppler resistive index and plasma creatinine and magnesium concentrations after surgical, unilateral ureteral obstruction in a rabbit model.

The aim of this study is to evaluate the changes in Doppler resistive index (RI) and plasma creatinine and magnesium concentrations after unilateral ureteral obstruction in a rabbit model. Fourteen adult female rabbits were used in this study. In seven rabbits, the left ureter was ligated with silk suture, and the control group was sham operated. Before surgery and on the second and seventh days after surgery, blood samples were obtained to measure plasma creatinine and magnesium concentrations. Doppler RIs of both kidneys were also measured before surgery and on the second and seventh days after the surgical procedure. With regard to magnesium levels, there was a significant within-subjects sessions difference [F(2, 20) = 15.21, P= 0.001] indicating a decrease through sessions. Magnesium concentrations decreased significantly at the postoperative second and seventh days compared to preoperative baseline levels (P= 0.003 and P= 0.001, respectively). Multifactorial analysis of variance was applied for each session separately with laterality, and groups as factors. The Doppler RI and the creatinine level did not show any significant differences or interactions for all sessions (P > 0.05). The decreasing plasma magnesium concentration after surgery may indicate ureteral injury; however, Doppler studies and creatinine levels may not be useful as well.

Genome size determination and coding capacity of Sodalis glossinidius, an enteric symbiont of tsetse flies, as revealed by hybridization to Escherichia coli gene arrays.

Recent molecular characterization of various microbial genomes has revealed differences in genome size and coding capacity between obligate symbionts and intracellular pathogens versus free-living organisms. Multiple symbiotic microorganisms have evolved with tsetse fly, the vector of African trypanosomes, over long evolutionary times. Although these symbionts are indispensable for tsetse fecundity, the biochemical and molecular basis of their functional significance is unknown. Here, we report on the genomic aspects of the secondary symbiont Sodalis glossinidius. The genome size of Sodalis is approximately 2 Mb. Its DNA is subject to extensive methylation and based on some of its conserved gene sequences has an A+T content of only 45%, compared to the typically AT-rich genomes of endosymbionts. Sodalis also harbors an extrachromosomal plasmid about 134 kb in size. We used a novel approach to gain insight into Sodalis genomic contents, i.e., hybridizing its DNA to macroarrays developed for Escherichia coli, a closely related enteric bacterium. In this analysis we detected 1,800 orthologous genes, corresponding to about 85% of the Sodalis genome. The Sodalis genome has apparently retained its genes for DNA replication, transcription, translation, transport, and the biosynthesis of amino acids, nucleic acids, vitamins, and cofactors. However, many genes involved in energy metabolism and carbon compound assimilation are apparently missing, which may indicate an adaptation to the energy sources available in the only nutrient of the tsetse host, blood. We present gene arrays as a rapid tool for comparative genomics in the absence of whole genome sequence to advance our understanding of closely related bacteria.

A novel application of gene arrays: Escherichia coli array provides insight into the biology of the obligate endosymbiont of tsetse flies.

Symbiotic associations with microorganisms are pivotal in many insects. Yet, the functional roles of obligate symbionts have been difficult to study because it has not been possible to cultivate these organisms in vitro. The medically important tsetse fly (Diptera: Glossinidae) relies on its obligate endosymbiont, Wigglesworthia glossinidia, a member of the Enterobacteriaceae, closely related to Escherichia coli, for fertility and possibly nutrition. We show here that the intracellular Wigglesworthia has a reduced genome size smaller than 770 kb. In an attempt to understand the composition of its genome, we used the gene arrays developed for E. coli. We were able to identify 650 orthologous genes in Wigglesworthia corresponding to approximately 85% of its genome. The arrays were also applied for expression analysis using Wigglesworthia cDNA and 61 gene products were detected, presumably coding for some of its most abundant products. Overall, genes involved in cell processes, DNA replication, transcription, and translation were found largely retained in the small genome of Wigglesworthia. In addition, genes coding for transport proteins, chaperones, biosynthesis of cofactors, and some amino acids were found to comprise a significant portion, suggesting an important role for these proteins in its symbiotic life. Based on its expression profile, we predict that Wigglesworthia may be a facultative anaerobic organism that utilizes ammonia as its major source of nitrogen. We present an application of E. coli gene arrays to obtain broad genome information for a closely related organism in the absence of complete genome sequence data.

Multi-site DNA polymorphism analyses of Leishmania isolates define their genotypes predicting clinical epidemiology of leishmaniasis in a specific region.

Leishmania isolates from 57 cases of human cutaneous (CL), human visceral (VL), and canine visceral (CVL) leishmaniasis in Turkey were grouped by multi-site DNA polymorphism analyses into five genotypes. The initial grouping was based on DNA heterogeneity of the faster-evolving mitochondrion (kinetoplast) minicircles and the intergenic regions of two nuclear repetitive genes. Taxonomic affiliation and phylogenetic relationships of the five genotypes were inferred by comparing them with reference species for sequence heterogeneity in a approximately 1.4 kb conserved single-copy gene, encoding N-acetylglucosamine-1-phosphate transferase (NAGT). Alignment of the available sequences revealed no gap, but up to 7% scattered base substitutions, suggesting that this functionally important gene is a suitable marker. Three genotypes are completely identical to the NAGTs of the reference species, identifying them as L. infantum, L. tropica. and L. major, respectively. The remaining two are recognized as L. major NAGT variants with one and four base substitutions, respectively. As expected, Maximum Likelihood analysis of the NAGT sequences separates them into three clades, corresponding to the three species. The majority of the isolates obtained are L. infantum and L. tropica, which have been known to cause infantile VL and anthroponotic CL in western and southeastern Turkey, respectively. Unexpected is the finding of Leishmania major variants and their dispersal, possibly as previously unrecognized clinico-epidemiologic entities of CL and VL.