Complete mitochondrial genome of the Amur hedgehog Erinaceus amurensis (Erinaceidae) and higher phylogeny of the family Erinaceidae.

We sequenced and characterized the complete mitogenome (KX964606) of the Amur hedgehog Erinaceus amurensis to provide more data for comparative mitogenomics of the genus Erinaceus (Erinaceidae). The mitogenome of E. amurensis is a circular molecule 16,941 bp long, consisting of a control region and a conserved set of 37 genes containing 13 protein-coding genes, 22 tRNA genes, and two rRNA genes (12S rRNA and 16S rRNA). The mitogenome of E. amurensis is AT-biased, with a nucleotide composition of 33.9% A, 21.1% C, 32.6% T, and 12.4% G. The mitogenomes of E. amurensis and the closely related hedgehog species E. europaeus, excluding the control region (66.7%), share over 90% sequence similarity. According to the inter-generic relationship based on six mitogenomes described from five genera of Erinaceidae, the subfamilies Erinaceinae and Galericinae are strongly supported as monophyletic groups, with each genus well placed within its own subfamily. Within the subfamily Erinaceinae, E. amurensis is a sister species to E. europaeus, and the relationship between Hemiechinus and Erinaceus is strongly supported. Within the subfamily Galericinae, the clade of Hylomys + Neotetracus was sister to that of Echinosorex, with clades supported by high values. Our findings will help to understand the codon usage pattern and molecular evolution of E. amurensis, and provide insight into inter-generic relationships within the family Erinaceidae. In future studies, the inclusion of mitogenomes from other genera would greatly enhance our understanding of higher phylogeny within the Erinaceidae.

Molecular detection by analysis of the 16S rRNA gene of fecal coliform bacteria from the two Korean Apodemus species (Apodemus agrarius and A. peninsulae).

Wild mouse feces can disseminate zoonotic microorganisms throughout a farm, which is a great threat to human health and can lead to economic loss through contaminated agricultural produce. To assess the microbial communities, especially fecal coliform bacteria, we used two methods. First, we isolated bacterial colonies onto the common media LB (lactose broth) agar, TSA (tryptic soy agar), and MRS (de Man, Rogosa, and Sharpe) agar, and then randomly select colonies from each plate and stocked them to the mother plate for genomic DNA isolation. Second, we analyzed bacterial colonies using the 16S rRNA gene molecular diagnostic method. Based on bacterial cultures and bacterial 16S rRNA gene markers, we detected four different bacterial species (Bacillus amyloliquefaciens, Escherichia coli, Staphylococcus xylosus, and Serratia liquefaciens) from fecal coliforms of the striped field mouse Apodemus agrarius and A. peninsulae in agricultural areas in South Korea. These results could help us to better understand the pathogen reservoirs of mice and initiate some preventive measures to mitigate the microbial risks associated with mouse fecal matter in agricultural production areas.

Effect of BRCA1 and XPG mutations on treatment response to trabectedin and pegylated liposomal doxorubicin in patients with advanced ovarian cancer: exploratory analysis of the phase 3 OVA-301 study.

BACKGROUND: We investigated the association of BRCA1 and XPG mutations with response rate (RR), progression-free survival (PFS) and overall survival (OS) in a subset of patients from a phase 3 clinical trial comparing the efficacy and safety of trabectedin + pegylated liposomal doxorubicin (PLD) versus PLD alone in patients with recurrent ovarian cancer. PATIENTS AND METHODS: A candidate array was designed based on the Breast Cancer Information Core database for BRCA mutation analyses. An exploratory analysis of BRCA1/XPG mutation status was conducted using a two-sided log-rank test and 0.05 significance in germline DNA samples from 264 women with failed first-line platinum-based chemotherapy, randomized (1 : 1) to trabectedin + PLD or PLD alone. RESULTS: Overall, 41 (16%) of the 264 women had BRCA1(mut) (trabectedin + PLD: n = 24/135, 18%; PLD: n = 17/129; 13%) and 17 (6%) had XPG(mut) (trabectedin + PLD: n = 8/135, 6%; PLD: n = 9/129, 7%). A higher RR was observed in BRCA1(mut) patients (20/41; 49%) versus BRCA1(wt) patients (62/223; 28%). Within the BRCA1(mut) group, trabectedin + PLD-treated patients had longer PFS and longer OS than PLD-treated patients (median PFS 13.5 versus 5.5 months, P = 0.0002; median OS 23.8 versus 12.5 months, P = 0.0086), whereas in BRCA1(wt) patients, OS was not significantly different (median OS: 19.1 versus 19.3 months; P = 0.9377). There were no differences in OS or PFS of patients with XPG(mut) between the two treatment arms. However, trabectedin + PLD-treated patients with XPG(mut) had a trend toward shorter PFS (median PFS: 1.9 versus 7.5 months; P = 0.1666) and OS (median OS: 14.5 versus 20.7 months; P = 0.1774) than those with XPG(wt). CONCLUSIONS: In this exploratory analysis, patients with recurrent ovarian cancer carrying the BRCA1(mut) had improved outcomes with trabectedin + PLD treatment compared with PLD alone. Prospective evaluation of BRCA status is likely an important evaluation for DNA-damaging agents and may significantly impact interpretation of clinical studies. XPG may be a biomarker of poor outcome in these patients.

Histomorphometric evaluation of maxillary molar roots and surrounding periodontium following molar intrusion in rats.

OBJECTIVES: To investigate periodontal tissue changes during and after molar intrusion in rats. SETTING AND SAMPLE POPULATION: The Department of Orthodontics at Yonsei University. Thirty 12-week-old male rats were assigned to 1 control and 5 experimental groups (n = 5 each). MATERIALS AND METHODS: In the experimental groups, two maxillary molars were intruded for 2 weeks; the control group underwent the same procedures without the intrusion force. After 2 weeks of intrusion, rats in one of the experimental groups and in the control group were killed. In the other four experimental groups, the new molar positions were either retained or not retained with an occlusal bite block for 1 or 2 weeks prior to euthanization. Histomorphometric analysis was performed for sulcus depth, osteoclast number per unit alveolar bone length, and root resorption area per unit root surface length. RESULTS: Sulcus depth increased during intrusion (P < 0.05), but decreased after 2 weeks of retention (P < 0.05). The number of osteoclasts increased during intrusion (P < 0.05), but subsequently decreased regardless of the retention regime (P < 0.05). Root resorption area increased after molar intrusion, irrespective of the retention regimen, relative to that of the control group (P < 0.05) and was the greatest after 2 weeks of retention. CONCLUSION: These results indicated that root resorption occurred during and after molar intrusion and that the surrounding periodontium remodeled accordingly as tooth positions were altered, regardless of retention regime.

Gene organization and characterization of the complete mitogenome of Hypsugo alaschanicus (Chiroptera: Vespertilionidae).

We sequenced and characterized the complete mitogenome of Hypsugo alaschanicus (Vespertilionidae) to provide more data for comparative mitogenomics of the genus Hypsugo. The mitogenome of H. alaschanicus is a circular molecule of 17,300 bp, consisting of a control region and a typically conserved set of 37 vertebrate genes containing 13 protein-coding genes (PCGs), 22 tRNA genes, and two rRNA genes (12S rRNA and 16S rRNA). The mitogenome of H. alaschanicus is AT-biased, with a nucleotide composition of 34.1 A, 30.9 T, 22.4 C, and 12.6% G. In the 13 mitochondrial PCGs of H. alaschanicus, the start codon ATG is used in all PCGs, except Nd2 and Nd3 (which use ATT), and Nd5 (which uses ATA). Eight PCGs (Nd1, Cox1, Cox2, Atp8, Atp6, Nd4L, Nd5, and Nd6) use TAA as the stop codon, while the stop codon AGA occurs only in Cytb. Incomplete stop codons (T--) are used in the other four PCGs (Cox3, Nd2, Nd3, and Nd4). These findings contribute to our understanding of the nucleotide composition and molecular evolution of the mitogenomes of the genus Hypsugo, and provide more data for comparative mitogenomics and higher phylogeny in the family Vespertilionidae.

Complete mitochondrial genome and codon usage of the Nepalese whiskered bat Myotis muricola (Vespertilionidae).

We sequenced and characterized the complete mitogenome of the Nepalese whiskered bat Myotis muricola (Vespertilionidae) to provide more data for comparative mitogenomics and codon usage in the genus Myotis (Vespertilionidae). The mitogenome of M. muricola is a circular molecule of 17,224 bp, consisting of a control region and a conserved set of 37 genes containing 13 protein-coding genes (PCGs), 22 tRNA genes, and two rRNA genes (12S rRNA and 16S rRNA). The mitogenome of M. muricola is AT-biased, with a nucleotide composition of 33.6% A, 29.7% T, 23.3% C, and 13.4% G. The total length of the 13 mitochondrial PCGs, excluding stop codons, is 11,376 bp, or 3792 amino acids. The relative synonymous codon usage (RSCU) of codons ending in A/T was generally higher than that for codons ending in G/C. The most frequently used codons are CTA(Leu) and CGA(Arg), with RSCU values greater than 2.0. The most rarely used codons, all terminating in G, are TCG(Ser), CCG(Pro), GCG(Ala), AAG(Lys), TGG(Try), CGG(Arg), and ACG(Thr), with RSCU values below 0.2. TCG(Ser) occurs only five times, and has the lowest RSCU value (0.091). These results are valuable for a better understanding of the molecular evolution of mitogenomes in the genus Myotis.

Detection and sequence analysis of the DNA repair gene RAD51 in the Korean spider Callobius koreanus (Amaurobiidae).

We identified a partial sequence (483 bp) of the RAD51 gene from the Korean spider Callobius koreanus. Sequence variation was found at one position during alignment with the human RAD51 gene sequence. This partial sequence included the region corresponding to exon 4 in the human RAD51 gene, which encodes 39 amino acids. These results show that the RAD51 gene is highly conserved between human and spiders.

Assessment of masticatory function in patients with non-sagittal occlusal discrepancies.

Non-sagittal occlusal discrepancies such as posterior cross-bite and anterior openbite are common types of malocclusion, but studies on masticatory function related to those malocclusions have been scarce. The aim of this study was to quantify the masticatory performance in patients with non-sagittal discrepancies compared to those with normal occlusion, using both objective and subjective measures. Maximum bite force and contact area using Dental Prescale(®) system as a static objective assessment, Mixing Ability Index (MAI) as a dynamic objective evaluation and food intake ability (FIA) as a subjective assessment were analysed from 21 people in normal occlusion (Group N) and 64 patients with posterior cross-bite (Group C), anterior openbite (Group O) or both (Group B). The differences of the maximum bite force, the contact area, the MAI and the FIA were compared, and their correlations were figured out. The non-sagittal malocclusion groups showed lower values in the maximum bite force, the contact area, the MAI and the FIA compared to those in the normal group (P < 0·0001). Compared to Group N, Groups C, O and B showed 61·5%, 42·1% and 40·1% of the maximum bite force, and 84%, 84% and 76% of hard food FIA, respectively. However, there were no significant differences among Groups C, O and B. The MAI showed higher correlation with the FIA (r = 0·38, P < 0·01), than with the maximum bite force and the contact area (both r = 0·24, P < 0·5). These results revealed that masticatory function in patients with non-sagittal discrepancies is significantly reduced both objectively and subjectively.

Use of permanent marker to deposit a protection layer against FIB damage in TEM specimen preparation.

Permanent marker deposition (PMD), which creates permanent writing on an object with a permanent marker, was investigated as a method to deposit a protection layer against focused ion beam damage. PMD is a simple, fast and cheap process. Further, PMD is excellent in filling in narrow and deep trenches, enabling damage-free observation of high aspect ratio structures with atomic resolution in transmission electron microscopy (TEM). The microstructure, composition, gap filling ability and planarization of the PMD layer were studied using dual beam focused ion beam, transmission electron microscopy, energy dispersive X-ray spectroscopy and electron energy loss spectroscopy. It was found that a PMD layer is basically an amorphous carbon structure, and that such a layer should be at least 65 nm thick to protect a surface against 30 keV focused ion beam damage. We suggest that such a PMD layer can be an excellent protection layer to maintain a pristine sample structure against focused ion beam damage during transmission electron microscopy specimen preparation.

Trabectedin plus pegylated liposomal doxorubicin in relapsed ovarian cancer: outcomes in the partially platinum-sensitive (platinum-free interval 6-12 months) subpopulation of OVA-301 phase III randomized trial.

BACKGROUND: OVA-301 is a large randomized trial that showed superiority of trabectedin plus pegylated liposomal doxorubicin (PLD) over PLD alone in relapsed ovarian cancer. The optimal management of patients with partially platinum-sensitive relapse [6-12 months platinum-free interval (PFI)] is unclear. PATIENTS AND METHODS: within OVA-301, we therefore now report on the outcomes for the 214 cases in this subgroup. RESULTS: Trabectedin/PLD resulted in a 35% risk reduction of disease progression (DP) or death [hazard ratio (HR) = 0.65, 95% confidence interval (CI), 0.45-0.92; P = 0.0152; median progression-free survival (PFS) 7.4 versus 5.5 months], and a significant 41% decrease in the risk of death (HR = 0.59; 95% CI, 0.43-0.82; P = 0.0015; median survival 23.0 versus 17.1 months). The safety of trabectedin/PLD in this subset mimicked that of the overall population. Similar proportions of patients received subsequent therapy in each arm (76% versus 77%), although patients in the trabectedin/PLD arm had a slightly lower proportion of further platinum (49% versus 55%). Importantly, patients in the trabectedin/PLD arm survived significantly longer after subsequent platinum (HR = 0.63; P = 0.0357; median 13.3 versus 9.8 months). CONCLUSION: This hypothesis-generating analysis demonstrates that superior benefits with trabectedin/PLD in terms of PFS and survival in the overall population appear particularly enhanced in patients with partially sensitive disease (PFI 6-12 months).

Trabectedin plus pegylated liposomal doxorubicin in relapsed ovarian cancer delays third-line chemotherapy and prolongs the platinum-free interval.

BACKGROUND: OVA-301 is a large randomized trial that showed superiority of trabectedin plus pegylated liposomal doxorubicin (PLD; CentoCor Ortho Biotech Products L.P., Raritan, NJ, USA). over single-agent PLD in 672 patients with relapsed ovarian cancer, particularly in the partially platinum-sensitive subgroup [platinum-free interval (PFI) of 6-12 months]. This superiority has been suggested to be due to the differential impact of subsequent (platinum) therapy. PATIENTS AND METHODS: a detailed analysis of subsequent therapies and survival outcomes in the overall population and in the subsets according to platinum sensitivity was therefore conducted. RESULTS: similar proportions of patients received subsequent therapy in each arm (76% versus 77%), including further platinum-based regimens (49% versus 55%). Patients in the trabectedin/PLD arm received subsequent chemotherapy at a later time (median delay 2.5 months versus PLD arm). Overall survival from subsequent platinum was significantly prolonged in the partially platinum-sensitive disease subset (hazard ratio = 0.63; P = 0.0357). CONCLUSION: the superiority of trabectedin/PLD over single-agent PLD in OVA-301 cannot be explained by differences in the extent or nature of subsequent therapies administered to these patients. On the other hand, these exploratory analyses support the hypothesis that the enhanced survival benefits in the partially platinum-sensitive subset might be due to an extended PFI leading to longer survival with subsequent platinum.

The role of psychological distress in laryngopharyngeal reflux patients: a prospective questionnaire study.

OBJECTIVES: To determine the role of psychological distress in laryngopharyngeal reflux patients and evaluate the correlation between symptoms, laryngeal signs, pH monitoring results and psychological profile. DESIGN: Prospective study. SETTING: Hanyang University Hospital, a university teaching hospital and tertiary referral center. PARTICIPANTS: One hundred and six patients who were diagnosed with laryngopharyngeal reflux by 24-h ambulatory double probe pH monitoring and 119 healthy controls visiting our health promotion center from January 2006 to June 2007. MAIN OUTCOME MEASURES: The psychological profile of laryngopharyngeal reflux patients measured by the Symptom Checklist-90-Revised questionnaire were evaluated and compared with those of healthy controls. The correlation between reflux symptom index, reflux finding score, parameters of pH monitoring and the Symptom Checklist-90-Revised profiles were also evaluated. RESULTS: On the Symptom Checklist-90-Revised questionnaire, the total mean T-scores of the nine symptom dimensions and three global indices of the laryngopharyngeal reflux patients were all below 50. The Global Severity Index, which indicates overall psychological distress, was normal in all of the patients. On comparison with the control group, no statistically significant difference was noted in the psychological profile except on the Somatisation scale where laryngopharyngeal reflux patients showed significantly higher scores. Reflux symptom index showed significant positive correlation with the number of reflux episodes, percentage of time which pH fell below 4 in total positions, and DeMeester score of the upper probe. The nine symptom dimensions and three global indices of Symptom Checklist-90-Revised questionnaire did not show any correlation with reflux symptom index, reflux finding score and the parameters of the 24-h ambulatory double probe pH monitoring. CONCLUSIONS: Laryngopharyngeal reflux patients did not demonstrate any significant level of psychological distress and their symptom severity showed significant positive correlation with reflux severity.

The effects of midazolam on the bispectral index after fetal expulsion in caesarean section under general anaesthesia with sevoflurane.

The effects of midazolam used with low concentration inhaled anaesthetics on the bispectral index (BIS) was investigated after fetal expulsion during caesarean section. Forty-five patients undergoing caesarean section received either normal saline (control, n = 15), or an intravenous bolus of 0.03 mg/kg (n = 15) or 0.05 mg/kg (n = 15) midazolam. Changes in BIS and maternal haemodynamics were monitored before induction, on intubation, at uterine incision, on delivery, at 3, 5 and 10 min after fetal expulsion, at subcutaneous tissue closure, at skin closure, on eye opening and at extubation. BIS values in the group that received 0.05 mg/kg midazolam were significantly lower than in the other two groups at 3, 5 and 10 min after fetal expulsion, and at subcutaneous tissue closure and skin closure. Values of BIS < 60 could only be maintained with 0.05 mg/kg midazolam and there was no delay in maternal emergence or recovery.

Hyperthermal neutral beam sources for material processing.

Hyperthermal neutral beams have a great potential for material processes, especially for etching and thin film deposition for semiconductor and display fabrication as well as deposition for various thin film applications. Plasma-induced damage during plasma etching is a serious problem for manufacturing deep submicron semiconductor devices and is expected to be a problem for future nanoscale devices. Thermal and plasma-induced damage is also problematic for thin film depositions such as transparent conductive oxide films on organic light emitting diodes or flexible displays due to high temperature processes in plasma environments. These problems can be overcome by damage-free and low-temperature processes with hyperthermal neutral beams. We will present the status of the hyperthermal neutral beam development and the applications, especially, in semiconductor and display fabrication and introduce potential applications of thin film growing for optoelectronic devices such as light emitting diodes.

Effect of nicardipine on haemodynamic and bispectral index changes following endotracheal intubation.

We investigated the effect of IV nicardipine on haemodynamic and bispectral index responses to the induction of general anaesthesia and intubation. Forty patients were randomly allocated to two groups of 20 to receive normal saline or nicardipine 15 microg/kg IV 30 s after induction. Ninety seconds later, tracheal intubation was performed. Systolic blood pressure, heart rate and bispectral index were measured at baseline, 1 min after induction, pre-intubation, and every minute until 5 min after endotracheal intubation. Rate-pressure product values were calculated. In the nicardipine group, systolic blood pressure decreased compared with the control group, and heart rate increased comparedwith the control group. Bispectral index and rate-pressure product showed no differences between the two groups. In conclusion, the administration of 15 microg/kg nicardipine IV does not affect anaesthetic depth in response to the induction of general anaesthesia and intubation.

Amiodarone offsets the cardioprotective effects of ischaemic preconditioning against ischaemia/reperfusion injury.

Both ischaemic preconditioning (IPC) and amiodarone protect against myocardial ischaemia. We examined whether a combination of IPC and amiodarone demonstrated an additive protective effect in isolated rat hearts (n = 40). The controls (group I) were subjected to ischaemia/ reperfusion injury; group II was subjected to cycles of IPC prior to ischaemia/ reperfusion injury; group III was subjected to ischaemia in the presence of amiodarone (10(-10) mol/1); and group IV was subjected to IPC followed by ischaemia in the presence of amiodarone (10(-10) mol/l). Amiodarone produced the best preserved left ventricular end-systolic pressure and dP/dtmax, less developed ventricular stiffness, the shortest arrhythmia duration, and the smallest infarct size among the groups. All of the myocardial protective effects against ischaemia/reperfusion injury were diminished or abolished when IPC and amiodarone were applied sequentially.

Redox factor-1: an extra-nuclear role in the regulation of endothelial oxidative stress and apoptosis.

The rac1 GTPase promotes oxidative stress through reactive oxygen species (ROS) production, whereas the DNA repair enzyme and transcriptional regulator redox factor-1 (ref-1) protects against cell death due to oxidative stimuli. However, the function of ref-1 in regulating intracellular oxidative stress, particularly that induced by rac1, has not been defined. We examined the role of ref-1 in vascular endothelial cell oxidative stress and apoptosis. Ref-1 was expressed in both the cytoplasm and nuclei of resting endothelial cells. Cytoplasmic ref-1 translocated to the nucleus with the oxidative trigger hypoxia/reoxygenation (H/R). Forced cytoplasmic overexpression of ref-1 suppressed H/R-induced oxidative stress (H(2)O(2) production), NF-kappaB activation, and apoptosis, and also mitigated rac1-regulated H(2)O(2) production and NF-kappaB transcriptional activity. We conclude that inhibition of oxidative stress is another mechanism by which ref-1 protects against apoptosis, and that this is achieved through modulation of cytoplasmic rac1-regulated ROS generation. This suggests a novel extra-nuclear function of ref-1.

Phase III double-blind placebo-controlled study of farnesyl transferase inhibitor R115777 in patients with refractory advanced colorectal cancer.

PURPOSE: To determine whether R115777 improves survival in patients with refractory advanced colorectal cancer (CRC) in a multicenter, double-blind, prospective randomized study. PATIENTS AND METHODS: Three hundred sixty-eight patients were randomly assigned to R115777 (300 mg twice daily) orally for 21 days every 28 days or placebo in a 2:1 ratio. All patients received best supportive care. The primary end point was overall survival; secondary end points were progression free survival, tumor response, toxicity, and quality of life. RESULTS: The two treatment groups were well balanced for baseline demographics, including previous chemotherapy for advanced CRC. The median overall survival for R115777 was 174 days (95% CI, 157 to 198 days), and 185 days (95% CI, 158 to 238 days) for those patients receiving placebo (P =.376). One patient achieved a partial response in the R115777 arm. Stable disease (> 3 months) was observed in 24.3% patients in the R115777 group compared to 12.8% in the placebo arm. This did not translate into a statistically significant increase in progression-free survival. Overall, treatment was well tolerated. There was an increased incidence of reversible myelosuppression (neutropenia, thrombocytopenia), rash, and grade 1 to 2 diarrhea in the R115777 arm. There was no statistically significant difference in quality of life between arms. CONCLUSION: Single agent R115777, given at this dose and schedule, has an acceptable toxicity profile, but does not improve overall survival compared to best supportive care alone in refractory advanced CRC.

Reduced cardiotoxicity and preserved antitumor efficacy of liposome-encapsulated doxorubicin and cyclophosphamide compared with conventional doxorubicin and cyclophosphamide in a randomized, multicenter trial of metastatic breast cancer.

PURPOSE: To determine whether Myocet (liposome-encapsulated doxorubicin; The Liposome Company, Elan Corporation, Princeton, NJ) in combination with cyclophosphamide significantly reduces doxorubicin cardiotoxicity while providing comparable antitumor efficacy in first-line treatment of metastatic breast cancer (MBC). PATIENTS AND METHODS: Two hundred ninety-seven patients with MBC and no prior chemotherapy for metastatic disease were randomized to receive either 60 mg/m(2) of Myocet (M) or conventional doxorubicin (A), in combination with 600 mg/m(2) of cyclophosphamide (C), every 3 weeks until disease progression or unacceptable toxicity. Cardiotoxicity was defined by reductions in left-ventricular ejection fraction, assessed by serial multigated radionuclide angiography scans, or congestive heart failure (CHF). Antitumor efficacy was assessed by objective tumor response rates (World Health Organization criteria), time to progression, and survival. RESULTS: Six percent of MC patients versus 21% (including five cases of CHF) of AC patients developed cardiotoxicity (P =.0002). Median cumulative doxorubicin dose at onset was more than 2,220 mg/m(2) for MC versus 480 mg/m(2) for AC (P =.0001, hazard ratio, 5.04). MC patients also experienced less grade 4 neutropenia. Antitumor efficacy of MC versus AC was comparable: objective response rates, 43% versus 43%; median time to progression, 5.1% versus 5.5 months; median time to treatment failure, 4.6 versus 4.4 months; and median survival, 19 versus 16 months. CONCLUSION: Myocet improves the therapeutic index of doxorubicin by significantly reducing cardiotoxicity and grade 4 neutropenia and provides comparable antitumor efficacy, when used in combination with cyclophosphamide as first-line therapy for MBC.

Experimental evolution of a dense cluster of residues in tyrosyl-tRNA synthetase: quantitative effects on activity, stability and dimerization.

A dense cluster of eight residues was identified at the crossing of two alpha-helices in tyrosyl-tRNA synthetase (TyrRS) from the thermophile Bacillus stearothermophilus. Its mechanism of evolution was characterized. Four residues of this cluster are not conserved in TyrRS from the mesophile Escherichia coli. The corresponding mutations were constructed in TyrRS(Delta1), a derivative of TyrRS from B. stearothermophilus in which the anticodon binding domain is deleted. Mutations I52L (i.e. Ile52 into Leu), M55L and L105V did not affect the activity of TyrRS(Delta1) in the pyrophosphate exchange reaction whereas T51P increased it. The kinetic stabilities of TyrRS(Delta1) and its mutant derivatives at 68.5 degreesC were determined from experiments of irreversible thermal precipitation. They were in the order L105V