BACKGROUND

Recent studies suggested an improved overall survival (OS) for BRCA2- versus BRCA1-associated epithelial ovarian cancer (EOC), whereas the impact of chemotherapy is not yet clear. In a nationwide cohort, we examined the results of primary treatment, progression-free survival (PFS), treatment-free interval (TFI), and OS of BRCA1 versus BRCA2 EOC patients.

METHODS

Two hundred and forty-five BRCA1- and 99 BRCA2-associated EOC patients were identified through all Dutch university hospitals. Analyses were carried out with the Pearson's Chi-square test, Kaplan-Meier, and Cox regression methods.

RESULTS

BRCA1 patients were younger at EOC diagnosis than BRCA2 patients (51 versus 55 years; P < 0.001), without differences regarding histology, tumor grade, and International Federation of Gynecology and Obstetrics (FIGO) stage. Complete response rates after primary treatment, including chemotherapy, did not differ between BRCA1 (86%) and BRCA2 patients (90%). BRCA1 versus BRCA2 patients had a shorter PFS (median 2.2 versus 3.9 years, respectively; P = 0.006), TFI (median 1.7 versus 2.8 years; P = 0.009), and OS (median 6.0 versus 9.7 years; P = 0.04). Differences could not be explained by age at diagnosis, FIGO stage or type of treatment.

CONCLUSIONS

PFS and OS were substantially longer in BRCA2- than in BRCA1-associated EOC patients. While response rates after primary treatment were similarly high in both groups, TFI, as surrogate for chemosensitivity, was significantly longer in BRCA2 patients.

The performance and reproducibility of the BoMED NCCOM3 thoracic electrical bioimpedance cardiograph (TEB) has been evaluated in volunteers and patients. In resting supine volunteers, we determined the coefficient of variability over short time periods (30 min) and over several days, and examined the effects of differences in electrode type and electrode placement. The mean (range) intra-subject coefficients of variation (CV) for thoracic fluid index (TFI) and stroke volume (SV) were 1.0% (0.4-1.8%) and 4.7% (2.1-8.5%), respectively over a 30-min period. The corresponding CV were 5.6% (2.3-10.9%) and 10.9% (6.1-14.8%) for measurements made at rest on four separate occasions. Use of different electrode types (RedDot and Medicotest) resulted in differences in TFI (P less than 0.01), but not in mean values for SV or cardiac output (Q); their use in individual subjects revealed differences of up to 20% in SV and Q. Alterations in electrode placement by 5 cm in the horizontal and diagonal planes produced no significant changes in TFI, SV or Q; changes in the longitudinal plane produced a graded change. Increases of 5 cm and 10 cm in thoracic length produced mean increases in TFI of 9.8% and 39.8%, respectively, and mean decreases in Q of 8.4% and 16.7% and SV of 7.5% and 15.8%. TEB measurements of Q and SV were compared with thermodilution (TD) in 16 intensive care patients. Mean (SEM) Q by TEB was 5.63 (1.10) litre min-1 compared with TD 4.38 (0.72) litre min-1 (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

During the genomics era, the use of thermostable DNA polymerases increased greatly. Many were identified and described-mainly of the genera Thermus, Thermococcus and Pyrococcus. Each polymerase has different features, resulting from origin and genetic modification. However, the rational choice of the adequate polymerase depends on the application itself. This review gives an overview of the most commonly used DNA polymerases used for PCR application: KOD, Pab (Isis™), Pfu, Pst (Deep Vent™), Pwo, Taq, Tbr, Tca, Tfi, Tfl, Tfu, Tgo, Tli (Vent™), Tma (UITma™), Tne, Tth and others.

The orphan receptor TR4, member of the nuclear hormone receptor family, is related to the orphan receptors TR2, COUP-TFI and ARP-1, and was originally cloned from the adult rat brain. The latter two orphan receptors have been implicated in central nervous system (CNS) development. To investigate a possible role for TR4 in brain development, expression of TR4 was studied in rat embryos. At embryonic days 14.5 and 19.5, high expression of TR4 was found in the CNS, while low expression was detected throughout the embryo. In postnatal rats, TR4 was mainly expressed in the hippocampus and cerebellum, resembling the expression pattern found in adult brain. These data show that like COUP-TFI and ARP-1, expression of TR4 becomes restricted to distinct areas. In adult brain, TR4 is predominantly expressed in granule cells of both hippocampus and cerebellum. The data suggest a possible role for TR4 during proliferation and maturation of brain structures.

BACKGROUND

The orphan receptors COUP-TF (chicken ovalbumin upstream promoter transcription factor) I and II are members of the nuclear receptor superfamily that play distinct and critical roles in vertebrate organogenesis. The involvement of COUP-TFs in cancer development has recently been suggested by several studies but remains poorly understood.

METHODS

MCF-7 breast cancer cells overexpressing COUP-TFI and human breast tumors were used to investigate the role of COUP-TFI in the regulation of CXCL12/CXCR4 signaling axis in relation to cell growth and migration. We used Immunofluorescence, western-blot, RT-PCR, Formaldehyde-assisted Isolation of Regulatory Elements (FAIRE) assays, as well as cell proliferation and migration assays.

RESULTS

Previously, we showed that COUP-TFI expression is enhanced in breast cancer compared to normal tissue. Here, we report that the CXCL12/CXCR4 signaling pathway, a crucial pathway in cell growth and migration, is an endogenous target of COUP-TFI in breast cancer cells. The overexpression of COUP-TFI in MCF-7 cells inhibits the expression of the chemokine CXCL12 and markedly enhances the expression of its receptor, CXCR4. Our results demonstrate that the modification of CXCL12/CXCR4 expression by COUP-TFI is mediated by the activation of epithelial growth factor (EGF) and the EGF receptor. Furthermore, we provide evidence that these effects of COUP-TFI increase the growth and motility of MCF-7 cells in response to CXCL12. Cell migration toward a CXCL12 gradient was inhibited by AMD3100, a specific antagonist of CXCR4, or in the presence of excess CXCL12 in the cell culture medium. The expression profiles of CXCR4, CXCR7, CXCL12, and COUP-TFI mRNA in 82 breast tumors and control non-tumor samples were measured using real-time PCR. CXCR4 expression was found to be significantly increased in the tumors and correlated with the tumor grade, whereas the expression of CXCL12 was significantly decreased in the tumors compared with the healthy samples. Significantly higher COUP-TFI mRNA expression was also detected in grade 1 tumors.

CONCLUSIONS

Together, our mechanistic in vitro assays and in vivo results suggest that a reduction in chemokine CXCL12 expression, with an enhancement of CXCR4 expression, provoked by COUP-TFI, could be associated with an increase in the invasive potential of breast cancer cells.

A dose-finding study was designed to determine the maximum tolerated dose (MTD) of a bimonthly 12-h (10:00 p.m to 10:00 a.m), timed flat infusion (TFI) of 5-fluorouracil (5-FU) plus irinotecan (CPT-11), without leucovorin (LV), for metastatic colorectal carcinoma (CRC). A total of 33 patients were treated. Seven dose levels included a fixed CPT-11 dose of 180 mg/m2 on days 1 and 15 (d(1,15)) and escalating doses of 5-FU 600-1200 mg/m2 on days 1-4 and 15-18 (d(1-4,15-18)). Dose-limiting toxicities (DLTs) were: grade 3-4 non-hematologic, grade 4 hematologic and any toxicity causing a more than a 2-week delay in treatment. The MTD was reached at the seventh dose level. DLTs were observed in 5/8 patients (63%): G3 diarrhea, 2 patients, associated with G3 mucositis in one instance; G4 neutropenia, 2 patients, associated with severe asthenia in 1 patient; G3 hand-foot syndrome, 1 patient. The recommended doses (RDs) were established at the sixth dose level: 5-FU, 1100 mg/m2/d(1-4,15-18); CPT-11 180 mg/m2/d(1,15) [5-FU and CPT-11 dose intensity (DI), 2200 and 90 mg/m2 per week (w), respectively]. At the recommended dose, the DLTs in 38 cycles were: mucositis, 2 cycles (5%); afebrile G4 neutropenia and hand-foot syndrome, 1 cycle (3%). In 24 assessable patients, the overall response rate was 37.5%. The present CPT-11/5-FU schedule is highly tolerable in an outpatient setting using the highest recommended 5-FU dose effective in advanced CRC.

OBJECTIVE

This study aimed to evaluate the clinical outcome of recurrent early-stage high-risk epithelial ovarian cancer patients.

METHODS

Demographic and clinicopathological data were collected from women enrolled in GOG 157 who underwent surgical staging and had recurrent disease. Survival probability was estimated using Kaplan-Meier method, and hazard ratio of death was analyzed using Cox regression model.

RESULTS

Of 74 women with recurrent early-stage high-risk ovarian cancer, the median age at recurrence was 63 years; 93% were White, 2.7% were Black, 2.7% were Asian, and 1.4% were Others. Fifty-eight percent had stage I, and the remainder had stage II disease. Clear cell, serous, endometrioid, mucinous, and other tumors consisted of 28.4%, 25.7%, 24.3%, 16.2%, and 5.4% of patients, respectively; in addition, 36.5% had ascites, 33.8% had positive cytology, and 43.2% had ruptured tumors. Fifty-eight percent underwent three cycles, and 42% had six cycles of adjuvant chemotherapy with paclitaxel and carboplatin. Recurrence was diagnosed clinically in 46% and radiographically in 54% of women. The median time from completion of primary chemotherapy to recurrence (treatment-free interval, <em>TFI</em>) was 21 months. Overall, median survival after recurrence was 24 months. Patients with longer (>24 months) <em>TFI</em> had a higher median survival after subsequent treatment at 35 months compared to only 10 months in those who recurred <or=24 months (p=0.003).

CONCLUSIONS

Although patients with primary early-stage high-risk ovarian cancer have an overall favorable prognosis, survival after recurrence is poor and comparable to those with recurrent advanced-stage disease. Novel therapeutic modalities are warranted in these high-risk patients.

To investigate the effect of fluorosis on the pattern and depth of etch of human enamel, 420 enamel specimens classified according to the Thylstrup and Fejerskov index (TFI) were etched with 37% phosphoric acid for varying times. The mean depths of etch for mildly fluorosed enamel (TFI = 1-3) were generally dependent on etching time (r = 0.55-0.76), and were not significantly different from the depth obtained for non-fluorosed (TFI = 0) specimens. The enamel specimens with more severe fluorosis (TF = 4) required longer etching time which showed little correlation with the mean depth of etch (r = 0.15-0.16). Furthermore, the etched specimens showed typical enamel etching patterns, independent of the severity of fluorosis. When the specimens with TFI = 4 were etched for 45 s, the subsurface organic network was evident, but this disappeared and typical etching patterns could be seen again when etching time was increased to 75-90 s. It is therefore concluded that the diagnosis of the severity of fluorosis must first be made whenever etching of fluorosed enamel is contemplated.

OBJECTIVE

To compare total fluid intake (TFI), defined as the sum of water and all other fluid types, assessed with a 24-h dietary (food and fluid) recall with mean TFI assessed with a 7-day fluid-specific record among adolescents and adults.

METHODS

This repeated cross-sectional study compared TFI as assessed by two fluid assessment instruments using a crossover approach. 290 adolescents (17.3 ± 0.8 years, 50 % boys) and 289 adults (43 ± 9.3 years, 50 % men) from Indonesia completed the study.

RESULTS

Significant correlations were observed between fluid intake assessed with the 24-h recall and the 7-day fluid record (r = 0.333; p < 0.001). The Bland-Altman method, however, showed an underestimation (bias) of mean TFI by a 24-h recall when compared with the 7-day fluid record [mean difference (95 % CI) -382 mL (-299, -465); p < 0.001]. The mean difference also increased with increasing TFI: Mean difference for the lowest and highest quartiles of TFI was 139 versus -1265 mL/day. The 7-day fluid record recorded two (95 % CI -1.9, -2.4; p < 0.0001) extra drinking acts compared with the 24-h recall, whereas the mean volume per drinking act was significantly higher with the 24-h recall [mean difference (95 % CI) 39 mL (31, 47); p < 0.001].

CONCLUSIONS

Compared with a 7-day fluid record, a 24-h dietary recall significantly underestimated TFI. Subjects recalled two less drinking acts, while estimating the volume consumed per drinking act to be larger. Since the adequate intakes for total water intake are based on median intakes observed in national surveys that most frequently used the 24-h recall method, they may potentially be underestimated.

It has been demonstrated that aqueous extract of Radix Hedysari Prescription and modified Radix Hedysari Prescription could improve the regeneration of injured peripheral nerve. Radix Hedysari is a main component in these two formulas. We hypothesized that Hedysari polysaccharides (HPS), a main active ingredient, could also enhance peripheral nerve regeneration after nerve injury in adult animals. In the present study, we examined the effects of HPS on sciatic nerve regeneration for 6 weeks following clamping in rats (administrated orally of 2 ml HPS liquid daily, 0.25 g/ml). The results showed that HPS was able to enhance sciatic function index (SFI) value, tibial function index (TFI) value, peroneal nerve function index (PFI) value, conduction velocity, and the number of regenerated myelinated nerve fibers, suggesting the potential clinical application of HPS for the treatment of peripheral nerve injury in humans.

OBJECTIVE

To evaluate the effects of fluorosis and bleaching on shear bond strengths of orthodontic brackets.

METHODS

A total of 45 (30 fluorosed and 15 non-fluorosed) non-carious freshly extracted human permanent premolar teeth which were extracted for orthodontic reasons and without any caries or visible defects were used in this study. Fluorosed teeth were selected according to the modified Thylstrup and Fejerskov index (TFI), which is based on the clinical changes in fluorosed teeth. First group consisted of 15 fluorosed teeth. Second group of fluorosed teeth were bleached with a 35% hydrogen peroxide office bleaching agent. Third group served as control. No bleaching procedure was applied. Orthodontic brackets were bonded with a light cure composite resin and cured with a halogen light. After bonding, shear bond strengths of the brackets were tested with Universal testing machine.

RESULTS

The results showed that fluorosis only and bleaching of fluorosed teeth significantly reduced the bond strengths of the orthodontic brackets (P < .001). Although bleaching of fluorosed teeth decreased the values more, no statistically significant difference was found between fluorosis group and fluorosis+bleaching group (P>> .05).

CONCLUSIONS

Fluorosis and bleaching of fluorosed teeth reduce bracket bond strength to enamel, but the bond strength with these still exceed the minimum 6 to 8 MPa required to expect adequate clinical performance.

OBJECTIVE

The aim was to examine stroke patients', carers' and volunteer supporters' experiences of peer support groups during hospital rehabilitation.

METHODS

Semi-structured interviews and questionnaires were analysed by inductive thematic analysis. Participants also answered a standardised Therapeutic Factors Inventory (TFI).

RESULTS

Five superordinate themes emerged for patients, carers and volunteer supporters. Three themes related to group processes; 'practical issues' (five subthemes), 'staff presence', 'similarity-difference', and comparison with other group members. 'Value of peers' (five subthemes) described beneficial outcomes. The 'similarity-difference' theme and four subthemes under 'value of peers' were related to items from the TFI which drew agreement from most participants. The supporters had some unique themes; two were concerned with group organisation, one was the experience of 'being helpful to others' and one described the experiences of training. As well as its links with themes, agreement with TFI items revealed experiences that did not emerge as themes; feeling secure, expressing emotions and increased independence.

CONCLUSIONS

Participation in the group was experienced as beneficial by participants. Benefits included helpful information, advice, making new connections and increased awareness of stroke. Participants identified important group processes such as upward and downward comparison. Responses to the TFI suggested that attendance brought therapeutic gains.

BACKGROUND

Previous research established significant relationships between total fluid intake (TFI) and urinary biomarkers of the hydration process in free-living males and females; however, the nature of this relationship is not known for pregnant (PREG) and lactating (LACT) women.

OBJECTIVE

To determine the relationship between urinary and hematological hydration biomarkers with TFI in PREG and LACT.

METHODS

Eighteen PREG/LACT (age: 31 ± 3 years, pre-pregnancy BMI: 24.26 ± 5.85 kg m-2) collected 24-h urine samples, recorded TFI, and provided a blood sample at 5 time points (15 ± 2, 26 ± 1, 37 ± 1 weeks gestation, 3 ± 1 and 9 ± 1 weeks postpartum during lactation); 18 pair-matched non-pregnant (NP), non-lactating (NL) women (age: 29 ± 4 years, BMI: 24.1 ± 3.7 kg m-2) provided samples at similar time intervals. Twenty-four-hour urine volume (U VOL), osmolality (U OSM), specific gravity (U SG), and color (U COL) were measured. Hematocrit, serum osmolality (S OSM), and serum total protein (S TP) were measured in blood.

RESULTS

Significant relationships were present between TFI and urinary biomarkers in all women (P < 0.004); these relationships were not different between PREG and NP, and LACT and NL, except U VOL in PREG (P = 0.0017). No significant relationships between TFI and hematological biomarkers existed (P>> 0.05).

CONCLUSIONS

Urinary biomarkers of hydration, but not hematological biomarkers, have a strong relationship with TFI in PREG, LACT, NP, and NL women. These data suggest that urinary biomarkers of hydration reflect TFI during pregnancy and breast-feeding.

Cytosolic phosphoenolpyruvate carboxykinase (EC 4.1.1.32; PEPCK-C) catalyzes the critical regulated step in adipocyte glyceroneogenesis. Numerous studies have shown that hormones and nutrients regulate PEPCK-C at the transcriptional level. We identified two upstream cis-acting DNA elements, gAF1/PCK1 and PCK2, that control adipocyte specific transcription of the PEPCK-C gene (Pck1). Both elements are direct repeat hexanucleotides separated by 1 bp (DR1 elements; variations of the sequence AGGTCAnAGGTCA). PCK2 is located 1 kbp upstream and is the essential element of an adipocyte specific enhancer. It is a peroxisome proliferator activated receptor gamma response element (PPRE) and directs the activation of the PEPCK-C gene during adipogenesis. In addition, it is a thiazolidinedione response element in mature adipocytes. In contrast, gAF1/PCK1, centered 445 bp upstream, is a pleiotropic element that mediates tissue specific glucocorticoid action-repression in adipocytes and induction in hepatocytes. It is a negative response element for PPARgamma, RXRalpha, COUP-TFII, and several unidentified proteins in some cell types, and a positive element for COUP-TFI and HNF4 in other cells type. The purpose of this presentation is to review the discovery and characterization of these two elements in adipocytes and describe how our work has contributed to understanding the mechanisms that control adipocyte glyceroneogenesis.

The aim of this study was to determine the effect of animal management and farm facilities on total feed intake (TFI), feed conversion ratio (FCR) and mortality rate (MORT) of grower-finishing pigs. In total, 310 batches from 244 grower-finishing farms, consisting of 454 855 Pietrain sired pigs in six Spanish pig companies were used. Data collection consisted of a survey on management practices (season of placement, split-sex by pens, number of pig origins, water source in the farm, initial or final BW) and facilities (floor, feeder, ventilation or number of animals placed) during 2008 and 2009. Results indicated that batches of pigs placed between January and March had higher TFI (P=0.006), FCR (P=0.005) and MORT (P=0.03) than those placed between July and September. Moreover, batches of pigs placed between April and June had lower MORT (P=0.003) than those placed between January and March. Batches which had split-sex pens had lower TFI (P=0.001) and better FCR (P<0.001) than those with mixed-sex in pens; pigs fed with a single-space feeder with incorporated drinker also had the lowest TFI (P<0.001) and best FCR (P<0.001) in comparison to single and multi-space feeders without a drinker. Pigs placed in pens with <50% slatted floors presented an improvement in FCR (P<0.05) than pens with 50% or more slatted floors. Batches filled with pigs from multiple origins had higher MORT (P<0.001) than those from a single origin. Pigs housed in barns that performed manual ventilation control presented higher MORT (P<0.001) in comparison to automatic ventilation. The regression analysis also indicated that pigs which entered to grower-finisher facilities with higher initial BW had lower MORT (P<0.05) and finally pigs which were sent to slaughterhouse with a higher final BW presented higher TFI (P<0.001). The variables selected for each dependent variable explained 61.9%, 24.8% and 20.4% of the total variability for TFI, FCR and MORT, respectively. This study indicates that farms can increase growth performance and reduce mortality by improving farm facilities and/or modifying management practices.

OBJECTIVE

The literature on the role of cytoreductive surgery beyond the secondary cytoreductive setting is limited. In this study, we reviewed the outcomes of patients with recurrent epithelial ovarian carcinoma who underwent tertiary cytoreduction.

METHODS

We performed a retrospective chart review of all patients with recurrent epithelial ovarian carcinoma who underwent tertiary cytoreduction at our institution from 1/1/90 to 12/31/02. Disease-specific survival (DSS) was calculated from the time of tertiary cytoreduction to last follow-up. Univariate and multivariate analyses were used to analyze outcomes and to identify potential prognostic factors.

RESULTS

A total of 26 patients were identified. The median follow-up after tertiary cytoreduction was 22.3 months (range, 0-71.7 months), with an overall median DSS of 33.4 months (95%CI, 20.4-46.4). On univariate analysis, treatment-free interval (TFI) before tertiary cytoreduction and residual disease after the procedure, as well as time to first recurrence, were found to be significant prognostic factors. Median DSS was 15 months for a TFI < or =12 months compared with 60.4 months for a TFI>> 12 months (P = 0.002). The median DSS for patients with residual disease < or =0.5 cm was 36.3 months compared with 10.6 months for patients with residual disease >0.5 cm (P <0.0001). On multivariate analysis, TFI and residual disease after tertiary cytoreduction retained prognostic significance (P < 0.05 for both).

CONCLUSIONS

Further cytoreductive surgery may offer a survival benefit in patients who experience a recurrence after secondary cytoreduction. This benefit appears to be greatest in patients with a longer TFI (>12 months) and in whom an optimal (< or = 0.5 cm) cytoreduction can be achieved.

Although an increasing number of nuclear orphan receptors have recently been identified, the number of known naturally occurring genes that are directly regulated by orphan receptors is still small. We have shown previously that the gene encoding the neuropeptide oxytocin (OT) is negatively regulated by the orphan receptors chicken ovalbumin upstream transcription factor I (COUP-TFI) and II. Here we show that the mouse OT gene promoter is activated by RORalpha, a representative of the ROR/RZR orphan receptor subfamily. Using promoter/chloramphenicol acetyltransferase reporter constructs in heterologous transfection assays, we determined that RORalpha action induces a <6-fold increase in promoter activity. By 5' and 3' deletion analysis, DNase footprint analysis and electrophoretic mobility shift assays, we found that RORalpha action is mediated by two 14 bp regions centered at 160 and 180 nucleotides upstream of the transcriptional initiation site. Both sites contain significant sequence identities with an established ROR recognition sequence. Mutations in either or both of these sites reduce significantly RORalpha-induced activation of the OT promoter. In view of the strong transcriptional activation exerted by RORalpha on the OT gene promoter and the widespread distribution of different members of the ROR/RZR family, interactions between ROR/RZR isoforms and the OT gene may form part of the multifactorial regulatory mechanisms that control OT gene expression in different tissues.

BACKGROUND

Infectious bursal disease (IBD) is a highly contagious immunosuppressive disease in young chickens caused by infectious bursal disease virus (IBDV). It causes huge economic losses to the poultry industry. The objective of this study is to develop a loop-mediated isothermal amplification (LAMP) method for the detection and discrimination of IBDV.

RESULTS

In this study, we applied reverse transcription loop-mediated isothermal amplification (RT-LAMP) to detect IBDV in one simple step and further identified the very virulent strain from non-vvIBDVs with a simply post-amplification restriction enzyme analysis. Based on sequence analysis, a set of two inner, two outer and two loop primers were designed to target the VP5 gene and they showed great specificity with no cross reaction to the other common avian pathogens. The detection limit determined by both color change inspection and agarose gel electrophoresis was 28 copies viral RNA, which was almost as sensitive as a real-time RT-PCR previous developed in our laboratory. We also identified a unique Tfi I restriction site located exclusively in non-vvIBDVs, so very virulent strain could be distinguished from current vaccine strains. By screening a panel of clinical specimens, results showed that this method is high feasible in clinical settings, and it obtained results 100% correlated with real-time RT-PCR.

CONCLUSIONS

RT-LAMP is a rapid, simple and sensitive assay. In combination with the Tfi I restriction analysis, this method holds great promises not only in laboratory detection and discrimination of IBDV but also in large scale field and clinical studies.

A temporal profile of lateral and fourth ventricle rat choroid plexus (LVCP and 4VCP, respectively) tissue injury and recovery was determined using alterations in K, Na, and H2O content and ultrastructure after 10 min of transient forebrain ischemia (TFI). At 0.5 h postischemia the LVCP displayed a maximum reduction in K content by 32% and a significant increase in Na content by 85% and H2O content by 22%. LVCP tissue K, Na, and H2O content returned to sham values by 24 h postischemia. Ultrastructural changes appeared more severe between 0.5 and 12 h postischemia, whereas by 24 h, normal ultrastructure was restored. Elevations in 4VCP tissue Na (P < 0.05) and H2O content, which were less than those in LVCP, gradually reached a maximum by 24 h compared with sham. No change in 4VCP tissue ultrastructure was observed. These results indicate that the LVCP tissue is more vulnerable than 4VCP in the bilateral carotid artery occlusion model but that it recovers in a timely manner after TFI. Furthermore, the ability of the LVCP tissue to rapidly recover suggests its functional importance in helping to restore brain homeostasis.

Transcriptional activation of silent genes can require the erasure of epigenetic marks such as DNA methylation at CpGs (cytosine-guanine dinucleotide). Active demethylation events have been observed, and associated processes are repeatedly suspected to involve DNA glycosylases such as mCpG binding domain protein 4, thymine DNA glycosylase (TDG), Demeter, and repressor of silencing 1. A complete characterization of the molecular mechanisms occurring in metazoan is nonetheless awaited. Here, we report that activation of the endogenous vitronectin gene in P19 cells by the nuclear receptor chicken ovalbumin upstream promoter-transcription factor I (COUP-TFI) is observed in parallel with the recruitment of TDG and p68 RNA helicase, two components of a putative demethylation complex. Interestingly, when activated, the vitronectin gene was loaded with DNA methyltransferases 3a and 3b (Dnmt3a/b), and a strand-biased decrease in CpG methylation was detected. Dnmt3a was further found to associate with COUP-TFI and TDG in vivo, and cotransfection experiments demonstrated that Dnmt3a/b can enhance COUP-TFI-mediated activation of a methylated reporter gene. These results suggest that Dnmt3a/b could cooperate with the orphan receptor COUP-TFI to regulate transcription of the vitronectin gene.

OBJECTIVE

Occlusive renovascular disease and hypertension may progress to CKD. Circulating levels of several biomarkers, including fibroblast growth factor (FGF)-23, Klotho, and soluble urokinase plasminogen activator receptor (suPAR), are altered in patients with CKD, but their role in essential hypertension (EH) and renovascular hypertension (RVH) remains unclear.

METHODS

Levels of FGF-23, Klotho, suPAR, plasminogen activator inhibitor (PAI)-1, tissue factor, and tissue factor pathway inhibitor (TFI) were measured in the inferior vena cava and renal vein of hypertensive patients with atherosclerotic renal artery stenosis (n=12) or age-matched participants with EH (n=12) and relatively preserved renal function. Single-kidney blood flow was measured to calculate renal release of markers. For control, peripheral vein levels were measured in healthy volunteers (HVs; n=12).

RESULTS

FGF-23 levels did not differ among the groups, whereas Klotho levels were lower in participants with RVH and EH than in HVs, and suPAR levels were elevated in patients with RVH compared with HVs and patients with EH (6.1±1.5 versus 4.4±1.9 and 3.2±1.2 ng/ml, P<0.05). PAI-1 levels were higher in patients with RVH than in patients with EH, but tissue factor and TFI levels were not statistically significantly different. After adjustment for GFR, Klotho levels remained decreased in both RVH and EH, and suPAR and PAI-1 levels remained elevated in RVH. eGFR correlated inversely with systemic and renal vein suPAR levels, and directly with systemic Klotho levels.

CONCLUSIONS

Klotho levels are low in hypertensive patients, whereas suPAR and PAI-1 levels are specifically elevated in RVH, correlating with GFR. Klotho, PAI-1, and suPAR may be markers of kidney injury in hypertensive patients.

The relationship between the utilization of 3H-thymidine in situ ([3H]-TdR fractional incorporation or TFI) and tumour growth delay after treatment with various cytotoxic agents has been examined. It is shown that (a) it is not possible to predict tumour growth delay, or to select the most effective agent, from changes in TFI 1 day after treatment; (b) there is a good correlation between tumour growth delay and the time for recovery of TFI to the pretreatment level; (c) there is a relationship within a tumour line between the depression of TFI 4 days after treatment and growth dealy induced by the same treatment. This relationship appears to be independent of the mechanism by which the agent exerts its cytotoxic effect.

In many tissues, the TF (Thomsen-Friedenreich) blood group antigen (Galbeta1-3GalNAc alpha-) behaves as an onco-foetal carbohydrate antigen, showing increased expression in malignancy and hyperplasia. Dietary lectins which bind the TF antigen have marked effects on proliferation of epithelial cells without cytotoxicity. This led us to speculate that anti-TF antibodies, including those that naturally occur in humans, might have similar effects. Five anti-TF antibodies, TF2 (human), TF5 (human), 5A8 (mouse), 8D8 (mouse) and BM22 (mouse), but not TFI (human) or 49H.9 (mouse), showed marked dose-dependent stimulation (95-192%) of [3H]thymidine incorporation by HT29 human colon cancer cells. Similar stimulation of proliferation of HT29 cells by these monoclonal antibodies (MAbs) was found when cell count assessment was used. Antibody-stimulated proliferation was inhibited by co-incubation with glycoproteins expressing Galbeta1-3GalNAc alpha- (asialo glycophorin or [Galbeta1-3GalNAc alpha-O-p-aminophenyl]n-human serum albumin). A proliferative effect of these antibodies was also demonstrated on human colon cancer cell lines LS174T and HT29-MTX but not on Caco-2 cells. Although immunoblotting showed similar binding patterns of all the antibodies on HT29 cell membrane extracts, there was little correlation between cell surface binding assessed by immunofluorescence and proliferative response, and internalization of the biotinylated antibody TF5 was demonstrated by confocal microscopy. Our results provide further evidence that cell surface glycoproteins which express TF antigen may play an important role in the regulation of cell proliferation and also suggest that human anti-TF antibodies may have proliferative effects on cells which express TF antigen.

OBJECTIVE

There is no standard treatment for recurrent epithelial ovarian cancer (EOC). As there are no curative options, many oncologists choose to treat women who recur with carboplatin, particularly if they are deemed to have platinum-sensitive disease. However, particularly in the era of platinum-taxane treatment as primary therapy, the utility of this treatment has not been established, nor is it clear whether the results of single-agent treatment are equivalent to that of combination therapy. We sought to determine the outcomes for patients with platinum-sensitive EOC who were treated with carboplatin-taxane therapy and received single-agent carboplatin (C) as second chemotherapy. In addition, we sought to compare these results to the outcomes in women who received carboplatin and paclitaxel (C + T) at first relapse.

METHODS

We identified 24 patients using our electronic institutional database with a histologically confirmed diagnosis of ovarian cancer that had a complete response to platinum-paclitaxel chemotherapy, relapsed greater than 6 months after treatment, and received single-agent carboplatin as second-line chemotherapy. We performed a subsequent comparison between a subgroup of this cohort and one that met the same inclusion criteria but received C + T at relapse between January 1998 and December 2000.

RESULTS

Eighteen patients were evaluable for response, and all were available for analysis of survival end points. For evaluable patients, the overall response rate was 39% (complete, 11%; partial, 28%). Twenty-two percent had stable disease. Six (25%) patients experienced a hypersensitivity reaction, including 1 who required hospitalization. The median overall survival was 22 months. The 2-year overall survival rate was 49%. Stratification by treatment-free interval (TFI) showed a 25% for a TFI between 6 and 12 months and 43% for a TFI>> 12 months. When a subgroup of these women (18/24) was compared to a cohort that received C + T (29), the combination was associated with a higher complete and overall response rate, 7 and 36% for C versus 45 and 71% for C + T (P = 0.02). The overall survival in women who received C was 26 months versus 42 months in the women who received C + T (P < 0.02).

CONCLUSIONS

Carboplatin as a single agent is effective therapy for recurrent ovarian cancer in women who recur following treatment with carboplatin and paclitaxel, and the treatment-free interval predicts response to single-agent carboplatin. However, our secondary analysis suggests that carboplatin and paclitaxel may produce a higher response rate and a survival benefit compared to C alone. This supports the conclusions of ICON4, which recently reported both overall and progression-free survival benefits with C + T over C in women with platinum-sensitive recurrent disease.