beta-Tubulin mutations are associated with resistance to 2-methoxyestradiol in MDA-MB-435 cancer cells

2-Methoxyestradiol is an estradiol metabolite with significant antiproliferative and antiangiogenic activity independent of estrogen receptor status. To identify a molecular basis for acquired 2-methoxyestradiol resistance, we generated a stable 2-methoxyestradiol-resistant (2ME2R) MDA-MB-435 human cancer cell line by stepwise exposure to increasing 2-methoxyestradiol concentrations. 2ME2R cells maintained in the presence of the drug and W435 cells maintained in the absence of the drug showed 32.34- to 40.07-fold resistance to 2-methoxyestradiol. Cross-resistance was observed to Vinca alkaloids, including vincristine, vinorelbine, and vinblastine (4.29- to 6.40-fold), but minimal resistance was seen to colchicine-binding agents including colchicine, colcemid, and AVE8062A (1.72- to 2.86-fold). No resistance was observed to paclitaxel and epothilone B, polymerizing agents (0.89- to 1.14-fold). Genomic sequencing identified two different heterozygous point mutations in the class I (M40) isotype of beta-tubulin at amino acids 197 (Dbeta197N) and 350 (Kbeta350N) in 2ME2R cells. Tandem mass spectrometry confirmed the presence of both wild-type and the mutant beta-tubulin in 2ME2R cells at the protein level. Consistently, treatment of parental P435 cells with 2-methoxyestradiol resulted in a dose-dependent depolymerization of microtubules, whereas 2ME2R cells remained unaffected. In contrast, paclitaxel affected both cell lines. In the absence of 2-methoxyestradiol, 2ME2R cells were characterized by an elevated level of detyrosination. Upon 2-methoxyestradiol treatment, levels of acetylated and detyrosinated tubulins decreased in P435 cells, while remaining constant in 2ME2R cells. These results, together with our structure-based modeling, show a tight correlation between the antitubulin and antiproliferative effects of 2-methoxyestradiol, consistent with acquired tubulin mutations contributing to 2-methoxyestradiol resistance.     

Primary squamous cell carcinoma of the stomach: a case report

Squamous cell carcinoma (SCC) originating from the stomach is a relatively rare entity. There are theories regarding the development of this rare tumor, but its exact pathogenesis remains obscure. Fewer than 100 cases of primary SCC of the stomach have been presented in the literature. Due to the advanced stage at the time of diagnosis in most of these cases, the prognosis is generally poor. In the case presented here, dissemination of the tumor to the transverse colon, gallbladder and omentum was present at diagnosis. Despite the tumor's advanced stage, complete remission was achieved after six courses of adjuvant chemotherapy with 5-flourouracil and cisplatin. No recurrence has been detected during follow-up. The patient has been healthy with no sign of the disease for three years.     

Cytarabine-induced fever complicating the clinical course of leukemia

The aim of this study is to assess the frequency and clinical characteristics of cytosine arabinoside-induced fever in patients with acute myeloid leukemia in remission, receiving high-dose (3 g/m2) consolidation therapy. We have investigated 77 consolidation cycles over a study period of 4 years. A strict definition of cytosine arabinoside-induced fever (i.e. patients without neutropenia and with negative blood cultures during the fever episodes) was used. Of the 77 consolidation cycles, fever due to cytarabine was detected in 33 cycles (43%). Median time of onset of fever from the beginning of first chemotherapy dose was 22 h and maximum temperature was in the range 38.0-39.7 degrees C (mean+/-SD: 38.8+/-0.5 degrees C). Median duration of fever was 10.15 h and did not exceed 72 h. There was no difference with regard to neutrophil and white blood cell counts between cycles with or without cytarabine fever. The cost of investigation of fever source was about US$2137. Our analysis suggests that 'cytarabine fever' is a frequent and often a self-limiting complication of high-dose cytosine arabinoside consolidation therapy, and cost-reductive approaches could be structured based on this background.     

The investigation of insulin resistance in patients with idiopathic hirsutism

Hirsutism, which is characterized by excessive growth of terminal hair in a male pattern, is a common clinical condition in women. It may result from various causes including polycystic ovary syndrome, nonclassic adrenal hyperplasia, adrenal or ovarian tumors, or it may be idiopathic. Idiopathic hirsutism (IH) is considered to be one of the most common forms of hirsutism. Although not universal, insulin resistance and hyperinsulinemia have been demonstrated in women with polycystic ovary syndrome. Because there are not enough data showing whether patients with IH also have insulin resistance, we intended to investigate the presence/absence of insulin resistance in women with IH. Thirty-two women with IH [mean age, 24.8 +/- 1.2 yr; body mass index (BMI), 24.6 +/- 0.8 kg/m2] and 17 healthy women (mean age, 25.8 +/- 0.6 yr; BMI, 22.5 +/- 0.6 kg/m2) were included in the study. Eight of 32 patients with IH had BMI higher than 30 kg/m2. The presence of insulin resistance was investigated by using basal insulin levels, the oral glucose tolerance test, the i.v. insulin tolerance test, and the homeostasis model assessment (HOMA) score in both groups. Six (18.7%) patients had impaired glucose tolerance (IGT). Overall, patients with IH had significantly (P < 0.05) higher basal insulin levels (10.5 +/- 1.1 mU/liter vs. 5.7 +/- 0.9 mU/liter) and HOMA scores (2.0 +/- 0.2 vs. 1.1 +/- 0.2) and lower plasma glucose disappearance rate values (5.2 +/- 0.2 vs. 6.0 +/- 0.3) than control subjects. However, patients with IGT were notably more obese than the patients with a normal glucose tolerance test. Analyses after omitting the patients with IGT showed that there was still a significant (P < 0.05) difference in terms of basal insulin levels and HOMA scores. Six of eight (75%) obese patients with IH showed IGT. These data suggest that IH is associated with insulin resistance and an increased prevalence of IGT in obese patients.     

Iron chelation with deferasirox in adult and pediatric patients with thalassemia major: efficacy and safety during 5 years' follow-up

Patients with β-thalassemia require lifelong iron chelation therapy from early childhood to prevent complications associated with transfusional iron overload. To evaluate long-term efficacy and safety of once-daily oral iron chelation with deferasirox, patients aged ≥ 2 years who completed a 1-year, phase 3, randomized trial entered a 4-year extension study, either continuing on deferasirox (deferasirox cohort) or switching from deferoxamine to deferasirox (crossover cohort). Of 555 patients who received ≥ 1 deferasirox dose, 66.8% completed the study; 43 patients (7.7%) discontinued because of adverse events. In patients with ≥ 4 years' deferasirox exposure who had liver biopsy, mean liver iron concentration significantly decreased by 7.8 ± 11.2 mg Fe/g dry weight (dw; n = 103; P < .001) and 3.1 ± 7.9 mg Fe/g dw (n = 68; P < .001) in the deferasirox and crossover cohorts, respectively. Median serum ferritin significantly decreased by 706 ng/mL (n = 196; P < .001) and 371 ng/mL (n = 147; P < .001), respectively, after ≥ 4 years' exposure. Investigator-assessed, drug-related adverse events, including increased blood creatinine (11.2%), abdominal pain (9.0%), and nausea (7.4%), were generally mild to moderate, transient, and reduced in frequency over time. No adverse effect was observed on pediatric growth or adolescent sexual development. This first prospective study of long-term deferasirox use in pediatric and adult patients with β-thalassemia suggests treatment for ≤ 5 years is generally well tolerated and effectively reduces iron burden. This trial was registered at www.clinicaltrials.gov as #NCT00171210.     

Is there an association between mastalgia and fibromyalgia? Comparing prevalence and symptom severity

To determine the prevalence of mastalgia in patients with fibromyalgia (FM) and the prevalence of FM in patients with mastalgia in order to investigate coexistence, and to compare the pain patterns in the case of mastalgia or FM alone versus the two in combination. Fifty consecutive patients with mastalgia and 50 consecutive patients with FM were assessed and examined both for the existence and severity of mastalgia and FM. A high proportion of patients with mastalgia (36%) fulfilled the criteria for FM and 42% had mastalgia in the FM group. Two distinctive entities mastalgia and FM, being both unexplained pain syndromes, seem to frequently coexist. Patients with mastalgia or FM should be thoroughly questioned considering each of the diseases so that in case of coexistence an appropriate therapy might be implemented for a successful pain management.     

Evaluation of QT dispersion in beta thalassaemia major patients

Cardiac complications are considered to be the primary cause of death in patients with beta thalassaemia major. QT dispersion is a marker variability of ventricular repolarization and is elevated in various high risk groups of patients. This study was carried out in patients with beta thalassaemia major to evaluate QT dispersion and to investigate the relationship between QT dispersion and body iron load. Sixty-two beta thalassaemia major patients were enrolled into the study. The average serum ferritin levels and liver iron concentration was assessed. For each patient, QT-QTc intervals and QT-QTc dispersions were calculated and V1S and V5R were measured. All the subjects underwent two-dimensional M mode echocardiogram and Doppler study. LVMI was found higher in thalassaemia major patients compared to control group. beta thalassaemia major patients showed significantly higher mean QT, QTc, QTd, and QTcd values compared to the control group. The mean V5R and V1S amplitudes were also higher in beta thalassaemia major patients. There was a positive correlation between LVMI and QTc, QTd and QTcd. However, there was no significant correlation between QT dispersion and serum ferritin and liver iron concentration. Prospective longitudinal studies are needed to assess the prognostic significance of these findings.