Differentiating mouse embryonic stem cells express markers of human endometrium.

BACKGROUND: Modeling early endometrial differentiation is a crucial step towards understanding the divergent pathways between normal and ectopic endometrial development as seen in endometriosis. METHODS: To investigate these pathways, mouse embryonic stem cells (mESCs) and embryoid bodies (EBs) were differentiated in standard EB medium (EBM). Immunofluorescence (IF) staining and reverse-transcription polymerase chain reaction (RT-PCR) were used to detect expression of human endometrial cell markers on differentiating cells, which were sorted into distinct populations using fluorescence-activated cell sorting (FACS). RESULTS: A subpopulation (50%) of early differentiating mESCs expressed both glandular (CD9) and stromal (CD13) markers of human endometrium, suggestive of a novel endometrial precursor cell population. We further isolated a small population of endometrial mesenchymal stem cells, CD45-/CD146+/PDGFR-ß+, from differentiating EBs, representing 0.7% of total cells. Finally, quantitative PCR demonstrated significantly amplified expression of transcription factors Hoxa10 and Foxa2 in CD13+ EBs isolated by FACS (p = 0.03). CONCLUSIONS: These findings demonstrate that mESCs have the capacity to express human endometrial cell markers and demonstrate potential differentiation pathways of endometrial precursor and mesenchymal stem cells, providing an in vitro system to model early endometrial tissue development. This model represents a key step in elucidating the mechanisms of ectopic endometrial tissue growth. Such a system could enable the development of strategies to prevent endometriosis and identify approaches for non-invasive monitoring of disease progression.

The role of oxidative stress in premalignant lesions.

PURPOSE OF INVESTIGATION: The authors aimed to evaluate serum total oxidant status (TOS), total antioxidant status (TAS), and oxida- tive stress index (OSI) in women with abnormal cervical cytology, to determine the association between serum oxidant and antioxidant status of these women, and the progression of abnormal cervical cytology. MATERIALS AND METHODS: A total of 75 women enrolled in the study: 20 women with a determination of atypical squamous cells of undetermined significance (ASCUS), 20 women with low squamous intraepithelial lesions (LSIL), 15 women with high squamous intraepithelial lesions (HSIL) and 20 healthy controls. Serum TOS and TAS were determined and OSI was calculated as the indicator of degree of oxidative stress. RESULTS: Serum TOS levels and OSI were highest in the HSIL group and there was a trend toward increasing serum TOS levels and OSI from ASCUS to HSIL group. CONCLUSION: The authors demonstrated that increased oxidative stress with altered antioxidant level is associated with abnormal cervical cytology. Serum oxidant and antioxidant status may provide guidance as a simple and cost-effective method for follow-up, treatment, and recommendation in all stages of lesions.

The early pregnancy volume measurements in predicting pregnancy outcome.

OBJECTIVE: The authors' aim was to develop a logistic regression model based on the ultrasonographic parameters on maternities which are showing a healthy improvement process during the first trimester of pregnancy. MATERIAL AND METHODS: Using 2D transvaginal ultrasound imaging, the crown rump length (CRL), yolk sac (YS), and gestational sac (GS) diameters were recorded in 225 women with gestational age < 11 weeks. Simplified V = 0.523 x length x height x width formula was used for the volume calculations. The results which ended in abortion were not included in the study. RESULTS: Linear regression analyses between yolk sac volume (YSV), YSV = 0.026 + 0.0018 x CRL (r²: 0.15; p < 0.001), gestational sac volume (GSV), GSV= -9.6 + 1.7 x CRL (r²: 0.52; p < 0.001), and embryo volume (EV), EV = -1.64 + 0.18 x CRL (r²: 0.4; p < 0.001), and CRL was made and a linear relationship was detected. The volume measurements showed a meaningful correlation with the week of pregnancy. The space in the GS (GS volume-embryo volume) increased as the age of pregnancy became older (r² = 0.46; p < 0.001). DISCUSSION: The first volume value was made in the first trimester by transvaginal ultrasonography, which showed a correlation with the age of pregnancy.

Does Pattern Scan Laser (PASCAL) photocoagulation really induce less VEGF expression in murine retina than conventional laser treatment?

To investigate the differences in the mRNA and protein expression levels of vascular endothelial growth factor (VEGF) in murine retina between mice subjected to conventional laser (AG) and those subjected to Pattern Scan Laser (PASCAL) system. Male C57BL/6 mice were randomly assigned to one of three groups: Group 1 (G1) receiving retinal scatter laser photocoagulation using with AG photocoagulator (n=16), Group 2 (G2) receiving retinal scatter laser photocoagulation using with PASCAL (n=16) and Group 3 (G3) served as an untreated control group (n=6). Molecular and morphological analyses of VEGF were performed on days 1, 2 and 5 by ELISA, real-time PCR and immuno-histochemical analysis. In samples which underwent AG (G1), when compared with the control group (G3), VEGF mRNA level increased 2.4 folds on day 2, whereas it decreased on day 5 (p□0.001). In samples which underwent PASCAL (G2), on the other hand, VEGF mRNA level increased 1.8 folds on day 1 and 2.2 folds on day 5 when compared with the control group (G3). In samples which underwent AG (G1), when compared with the control group (G3), VEGF protein level increased significantly on day 2, whereas it decreased on day 5 (p□0.001). In group G2, the VEGF levels in the sensory retina significantly increased as compared to control groups at both 2 and 5 days after laser photocoagulation using PASCAL laser (p=0.012, both time points). The peak expressions of VEGF protein in samples which underwent PASCAL and conventional laser were found on day 5 and day 2 respectively. In retinas of PASCAL-treated mice, VEGF immunoreactivity gradually increased during the 5-day follow-up. However, in argon laser group, the strongest VEGF immunoreactivity was detected on day 2, then started to decrease on day 5. In summary, the expression of VEGF protein and mRNA gradually increase during a 5-day follow-up period in PASCAL-treated mouse eyes, whereas in AG group they reach their peak levels on the second day of follow-up and started decreasing after then. These results may also explain why the PASCAL system is less effective in regressing neovascularization in the clinic.

No additive effects of inhaled iloprost and prone positioning on pulmonary hypertension and oxygenation in acute respiratory distress syndrome.

BACKGROUND: In acute respiratory distress syndrome (ARDS), pulmonary hypertension is associated with a poor prognosis. Prone position is effective to improve oxygenation whereas inhaled iloprost can treat pulmonary hypertension. However, combination of these interventions has not been examined before. The hypothesis was that this combination had additive effects on oxygenation and pulmonary hemodynamics as compared with each intervention alone. METHODS: In a prospective, randomized cross-over study, ten pigs were anesthetized, intubated and ventilated with volume controlled ventilation. Carotid, jugular venous and pulmonary artery catheters were inserted. ARDS was induced with oleic acid (0.20 mL/kg). Measurements were repeated in randomized different sequences of prone or supine positions with or without iloprost inhalation (220 ng/kg/min) (four combinations). Systemic and pulmonary arterial pressures; arterial and mixed venous blood gases; and Qs/Qt and the resistances were recorded. RESULTS: Iloprost decreased pulmonary artery pressures (for MPAP: P=0.034) in both supine (37±10 vs. 31±8 mmHg; P<0.05) and prone positions (38±9 vs. 29±8 mmHg; P<0.05); but did not obtain a significant improvement in oxygenation in both positions. Prone position improved the oxygenation (p<0.0001) compared to supine position in both with (361±140 vs. 183±158 mmHg, P<0.05) or without iloprost application (331±112 vs. 167±117 mmHg, P<0.05); but did not achieve a significant decrease in MPAP. CONCLUSION: Although iloprost reduced pulmonary arterial pressures, and prone positioning improved oxygenation; there are no additive effects of the combination of both interventions on both parameters. To treat both pulmonary hypertension and hypoxemia, application of iloprost in prone position is suggested.

A recruitment maneuver improves apneic oxygenation.

BACKGROUND: Apneic oxygenation (AO) is applied during surgery and in intensive care units. Even with AO, apnea is associated with progressive hypoxemia, limiting the tolerable amount of time in AO. This experimental study evaluates the effects of a recruitment maneuver (RM) on oxygenation, CO2 retention, and survival times during prolonged apnea, supported or not supported with intratracheal apneic oxygenation. METHODS: Following Ethic Committee approval, 15 male Sprague-Dawley rats were anesthetized and ventilated with PCV and FiO2:1 for 15 minutes. After obtaining a basal arterial blood-gas sample, the rats were randomized into 3 groups and disconnected from the ventilator: group (G) 1 (N.=6): AO with a cannula inserted into the carina; G2 (N=6): RM (40 cm H2O CPAP applied for 30 seconds) before AO; and G3 (N.=3): no application after disconnection (G3 was stopped after the first 3 subjects died within 3 minutes). Further arterial blood-gas samples were taken after 1, 3, and 6 minutes (T1, T3, and T6). Survival times after the start of AO were recorded. RESULTS: G2 was associated with significantly higher values of PaO2 at T3 and T6 when compared to G1 (345±56 vs. 233±65 mm Hg at T3 and 258±31 vs. 180±31 mm Hg at T6, respectively, P<0.05). There were significant changes in PaO2, pH, and PaCO2 over time in all subjects, but no differences were observed between G1 and G2 in pH or PaCO2. Survival time in G2 was significantly longer as opposed to G1 (G1: 10.3±2.3 min; G2: 14.3±3.6 min; P<0.05). CONCLUSION: RM prior to AO prolongs tolerance to apnea, probably by increasing the time before intolerable hypoxemia occurs, without a significant difference in PaCO2 levels.

Application of struvite precipitation coupled with biological treatment to slaughterhouse wastewaters.

In this study, struvite precipitation coupled with an activated sludge process was applied to slaughterhouse wastewaters. Biological treatability characteristics of the wastewater were evaluated in a wide organic loading range of 0.06-0.42 g COD (g MLVSS)(-1)d(-1) to assess COD removal as well as the extent of nitrification. Results of biological treatment of raw wastewater indicated that COD removal varied between 88% and 99% and complete nitrification was achieved at 0.1 g COD (g MLVSS)(-1)d(-1) and lower organic loadings. Biological treatment of the struvite-precipitated sample, which required no nitrification, yielded 86% COD removal at the organic removal of 0.3 g COD (g MLVSS)(-1)d(-1). Struvite precipitation of both raw and biologically treated wastewater was at effective stoichiometric magnesium and phosphate doses to ammonia and between pH values of 9.0 and 9.5, providing ammonia concentrations of 20-30 mg N L(-1) in the effluent and being independent of initial concentrations. The application of struvite precipitation both prior to and after biological treatment resulted in similar effluent qualities and provided the additional benefit of having a high-rate activated sludge system instead of a low organic loading system with nitrification-denitrification. An additional benefit of struvite precipitation was the production of sludge which had recovery potential as a fertilizer. Among the tested treatment schemes, biological treatment following struvite precipitation seemed to be more advantageous in terms of process stability.

Effects of thoracic epidural anaesthesia on pulmonary venous admixture and oxygenation during one-lung ventilation.

BACKGROUND: In this clinical randomized study, the effects of four anaesthesia techniques during one-lung ventilation [total intravenous anesthesia (TIVA) with or without thoracic epidural anaesthesia (TEA) (G-TIVA-TEA and G-TIVA), isoflurane anaesthesia with or without TEA (G-ISO-TEA and G-ISO)] on pulmonary venous admixture (Qs/Qt) and oxygenation (OLV) were investigated. METHODS: In 100 patients (four groups, 25 patients in each) undergoing thoracotomy, a thoracic epidural catheter was inserted pre-operatively. In G-TIVA-TEA and G-ISO-TEA, bupivacaine 0.1% + 0.1 mg/ml morphine was administered intra-operatively (10 ml of first bolus + 7 ml/h infusion). Propofol infusion or isoflurane concentration was adjusted to keep a bispectral index (BIS) of between 40 and 50 in all groups. FiO(2) was 0.8 during OLV and 0.5 before and after OLV. Partial arterial and central venous oxygen pressures (PaO(2) and PvO(2)), arterial and venous oxygen saturations and Qs/Qt values were recorded before, during and after OLV. RESULTS: During OLV, PaO(2) was significantly higher and Qs/QT significantly lower in G-TIVA-TEA and G-TIVA compared with G-ISO-TEA and G-ISO (PaO2: 188 +/- 36; 201 +/- 39; 159 +/- 33; 173 +/- 42 mmHg, respectively; Qs/Qt: 31.2 +/- 7.4; 28.2 +/- 7; 36.7 +/- 7.1; 33.7 +/- 7.7%, respectively). No statistical changes were observed in patients with TEA compared with without TEA in any measurement. CONCLUSION: During OLV, TEA does not significantly affect the oxygenation and Qs/Qt and can be used safely regardless of whether TIVA or inhalation techniques are used.

PER-1- and OXA-10-like beta-lactamases in ceftazidime-resistant Pseudomonas aeruginosa isolates from intensive care unit patients in Istanbul, Turkey.

The presence of PER-1- and OXA-10-like beta-lactamases was investigated by PCR in 49 ceftazidime-resistant Pseudomonas aeruginosa isolates from patients hospitalised in the 24-bed general intensive care unit of the Istanbul Faculty of Medicine during a 12-month period between February 1999 and February 2000. The clonal relatedness of the isolates was investigated by random amplified polymorphic DNA (RAPD) analysis, and the sequences of the PER-1 and OXA genes from all isolates were determined. The rates of resistance of the isolates to imipenem, aztreonam and cefepime were 98%, 92% and 96%, respectively, and to piperacillin and piperacillin-tazobactam were 41% and 37%, respectively. Using the double-disk synergy test, 37% (18/49) of the isolates were identified as extended-spectrum beta-lactamase producers. The PER-1 gene was identified in 86% (42/49) and the OXA-10 gene in 55% (27/49) of the ceftazidime-resistant isolates. Of isolates carrying the PER-1 gene, 48% (20/42) also carried the OXA-10 gene. The respective nucleotide sequences were identical for each isolate. Sixteen RAPD patterns were detected among the PER-1-positive isolates, but 60% (25/42) of the PER-1-positive isolates belonged to two distinct patterns, while the remainder exhibited a wide clonal diversity. The results indicated that the prevalence of PER-1- and OXA-10-like beta-lactamases remains high among ceftazidime-resistant P. aeruginosa isolates in Turkey.

Solitary rib lesions in bone scans of patients with breast carcinoma.

The aetiology of solitary rib lesions detected on bone scans was evaluated retrospectively. Seventy-five patients with breast carcinoma, and each with a solitary hot spot on a bone scan, were included in the study. The aetiology of the solitary rib lesions was determined by using all available clinical, laboratory and radiological data, and was clarified in 65 of the 75 patients, and not clarified in the remaining 10. In 17 of those 65 (26.1%), the aetiology of increased uptake was malignant; while in 48 of the 65 (73.8%) it was benign in origin. Linear lesions were mainly metastatic in origin (seven of nine, 77.7%) whereas focal lesions were mostly benign in origin (46 of 56, 82.19%). In the group of 16 hot spots located at the anterior rib end, 14 (87.5%) were benign, and two (12.5%) were malignant in origin. The relation between mastectomy side and the distribution of anterior and anterior rib end localization of hot spots was also investigated. Twenty-three of 32 rib lesions (71.8%) were on the same side as the surgery. In conclusion, solitary rib lesions on bone scans in patients with breast carcinoma are frequently benign in origin, especially if they are focal and located at the anterior rib end.

Reliability of procalcitonin as a severity marker in critically ill patients with inflammatory response.

Procalcitonin (PCT) is increasingly recognised as an important diagnostic parameter in clinical evaluation of the critically ill. This prospective study was designed to investigate PCT as a diagnostic marker of infection in critically ill patients with sepsis. Eighty-five adult ICU patients were studied. Four groups were defined on the basis of clinical, laboratory and bacteriologic findings as systemic inflammatory response syndrome (SIRS) (n = 10), sepsis (n = 16), severe sepsis (n = 18) and septic shock (n = 41). Data were collected including C-reactive protein (CRP), PCT levels and Sequential Organ Failure Assessment and Acute Physiology and Chronic Health Evaluation II scores on each ICU day. PCT levels were significantly higher in patients with severe sepsis and septic shock (19.25 +/- 43.08 and 37.15 +/- 61.39 ng/ml) than patients with SIRS (0.73 +/- 1.37 ng/ml) (P < 0.05 for each comparison). As compared with SIRS patients, plasma PCT levels were significantly higher in infected patients (21.9 +/- 47.8 ng/ml), regardless of the degree of sepsis (P < 0.001). PCT showed a higher sensitivity (73% versus 35%) and specificity (83% versus 42%) compared to CRP in identifying infection as a cause of the inflammatory response. Best cut-off levels were 1.31 ng/ml for PCT and 13.9 mg/dl for CRP. We suggest that PCT is a more reliable marker than CRP in defining infection as a cause of systemic inflammatory response.

[The effect of pentaglobin therapy on prognosis in patients with severe sepsis]. / Agir sepsis hastalarinda Pentaglobin tedavisinin prognoz üzerine etkisi.

This study was designed to assess the effects of polyclonal immunoglobulin administration on septic shock incidence and prognosis in patients with severe sepsis. Patients with severe sepsis were randomly allocated into two groups. One group (n = 21) received 5 ml/kg/day IgM enriched immunoglobulin preparation (Pentaglobin) for 3 days. Other group did not receive immunoglobulins (n = 18). Simplified Organ Failure Assessment (SOFA) scores, leucocyte count, duration of mechanical ventilation, ICU stay, duration of severe sepsis did not show significant differences between the groups, as regards to septic shock incidence and mortality. However, a significant decrease in procalcitonin levels were detected only in patients who received pentaglobin (p = 0.001). Mortality rate was 5/21 (23.8%) in pentaglobin group and 5/18 (27.7%) in the control group. Although pentaglobin therapy could not achieve a statistically significant improvement in septic shock occurrence and mortality, the constant reduction in procalcitonin levels indicated the beneficial effects of immunotherapy on the severity of inflammatory response to infection in severe sepsis.

[Percutaneous endoscopic gastrostomy in the ICU]. / Yogun bakim ünitesinde perkutan endoskopik gastrostomi.

Aspiration pneumonia due to gastroesophageal reflux is a frequent complication in ICU. The most commonly chosen method for long-term enteral access is gastrostomy and this method also reduces the risk of aspiration and shortens the hospital stay. We evaluated 31 patients in whom PEG was performed between 1997-98 in our unit. Indication of PEG was long-term ICU stay and coma which necessitate long term enteral nutrition. Mean age of our patients were 60.5((19.4). We evaluated the aspiration by clinical inspection. Aspiration and interruption of feeding was observed 1.57 ((1.43) times before PEG and 0.67 ((0.73) times after PEG. Further more positive tracheal aspirate culture were seen 3.14 ((1.95) times before PEG and 1.52 ((1.47) times after PEG.