Recent developments of capillary electrophoresis in seawater analysis.

Recent applications of capillary electrophoresis (CE) to the determination of various seawater analytes are critically examined, with the aim to reveal method's state-of-the-art and possible future research trends in the area. Given highly developed separation and detection methodology, emphasis is given to the most advantageous sample preconcentration strategies used to evolve the method's practical utility, particularly to low-level analytes. Analytical performance of CE applied to seawater analysis is illustrated with a selection of real-world applications published from 2006, from which it appears that the primary developmental tendency is presently confined to a transit from inorganic to organic seawater analytes.

Proportion of membrane proteins in proteomes of 15 single-cell organisms analyzed by the SOSUI prediction system.

A software system, SOSUI, was previously developed for discriminating between soluble and membrane proteins and predicting transmembrane regions (Hirokawa et al., Bioinformatics, 14 (1998) 378-379). The performance of the system was 99% for the discrimination between two types of proteins and 96% for the prediction of transmembrane helices. When all of the amino acid sequences from 15 single-cell organisms were analyzed by SOSUI, the proportion of predicted polytopic membrane proteins showed an almost constant value of 15-20%, irrespective of the total genome size. However, single-cell organisms appeared to be categorized in terms of the preference of the number of transmembrane segments: species with small genomes were characterized by a significant peak at a helix number of approximately six or seven; species with large genomes showed a peak at 10 or 11 helices; and species with intermediate genome sizes showed a monotonous decrease of the population of membrane proteins against the number of transmembrane helices.

Standardization of capillary zone electropherograms obtained by using field enhanced sample stacking.

Migration times in a capillary zone electropherogram obtained by using the field enhanced sample stacking technique are strongly affected by the injected sample volume. That is, the migration times significantly decrease with the increase of the sample volume. To avoid inaccurate qualitative analysis due to the above phenomena, the time axis of the electropherograms was converted into an effective mobility axis using our conversion method taking account of the temperature increase in the separation tube and relaxation of the potential gradient of the separation field. After the conversion, accurate qualitative analysis was possible in spite of drastic change of the migration time, suggesting our conversion method could be successfully used for the standardization of electropherograms obtained even by using the stacking effect. The cause of the decrease of the migration time in the stacking process was briefly discussed.

New method for standardization of electropherograms obtained in capillary zone electrophoresis.

A new method for standardization of electropherograms obtained by capillary zone electrophoresis was proposed, where the migration time axis was replaced by the effective mobility axis. The mobility increase due to temperature increase by Joule heating and the relaxation effect of the potential gradient were eliminated successfully by introducing a temperature coefficient for mobility expression and a delay time, respectively. The precision of the mobility evaluated by the proposed conversion methods was evaluated for a model sample. By using the conversion method, almost the same electropherograms could be obtained even from the electropherograms originally obtained by using different hardware conditions.

High-sensitivity capillary gel electrophoretic analysis of DNA fragments on an electrophoresis microchip using electrokinetic injection with transient isotachophoretic preconcentration.

The research adopted a single-channel microchip as the probe, and focused electrokinetic injection combined with transient isotachophoresis preconcentration technique on capillary electrophoresis microchip to improve the analytical sensitivity of DNA fragments. The channel length, channel width and channel depth of the used microchip were 40.5 mm, and 110 and 50 microm, respectively. The separation was detected by CCD (charge-coupled device) (effective length=25 mm, 260 nm). A 1/100 diluted sample (0.2 mg/l of each DNA fragment) of commercially available stepladder DNA sample could be baseline separated in 120 s with S/N=2-5. Compared with conventional chip gel electrophoresis, the proposed method is ideally suited to improve the sensitivity of DNA analysis by chip electrophoresis.

Isotachophoretic separation behavior of rare-earth EDTA chelates and analysis of minor rare-earth elements in an iron ore by bidirectional isotachophoresis-particle-induced X-ray emission.

Mobilities of 16 anions of rare-earth-EDTA 1:1 chelate (RE-EDTAs) were isotachophoretically measured by using two leading electrolytes (pH 3.6 and 6.0) in order to assess their separation behavior. The leading electrolyte was 20 mM hydrochloric acid. The pH of the solution was adjusted to 3.6 by adding beta-alanine and to 6.0 by adding histidine. The obtained mobilities were very close to each other in the range 20.1x10(-5)-21.9x10(-5) cm2 V(-1) s(-1) with the minimum mobilities for Pr-EDTA and Nd-EDTA for pH 3.6 and 6.0, respectively, and pH dependence was hardly observed. On the basis of the above knowledge. minor rare-earth elements in a standard iron ore sample were determined as RE-EDTAs by bidirectional isotachophoresis-particle-induced X-ray emission (PIXE), where the Fe(II) matrix digested by alkali fusion was separated as Fe(II)Phen3(2+) (phen = 1,10-phenanthroline). Since 5% of the total iron was still detected as Fe(III)EDTA- and might disturb PIXE analysis of RE-EDTA-, itaconic acid was used as the spacer for Fe(III)EDTA- and RE-EDTA-. The fractions of RE-EDTA- were successfully analyzed off-line by a multielemental analytical method, PIXE [analytical result (3.62% (w/w) as RE2O3]; the nominal value was 3.37% (w/w) as RExOy.

Large vessel arteritis associated with human herpesvirus 6 infections.

A 9-month-old boy presented with chronic arteritis of the aorta and its major branches. The clinical manifestations at onset of his illness were compatible with those of Kawasaki syndrome. However, the febrile period lasted for 2 months despite various immunosuppressive therapies, and the levels of C-reactive protein remain high 18 months after onset. Elevated circulating immune complexes, decreased serum complement levels, hypergammaglobulinaemia and monoclonal gammopathy were observed. Active HHV-6 infection was shown by increased serum levels of antihuman herpesvirus-6 (HHV-6) IgG and IgM antibodies, and positive HHV-6 DNA in sera, peripheral blood mononuclear cells (PBMNC) and lymph nodes. HHV-6 was actively replicating in PBMNC and lymph nodes, as shown by the detection of transcripts for the virus structural antigen. These results suggest that large vessel arteritis can be associated with HHV-6 infection.

Intraarterial infusion of 4'-O-tetrahydropyranyl-adriamycin for head and neck tumors.

THP-adriamycin was administered in a dose of 5 to 20 mg, 3 times a week, and the results showed 4 cases of CR, 5 cases of PR, 3 cases of MR and 2 cases of NC with the accumulated total dose of less than 100 mg. Histological effects were slight in a total dose of less than 50 mg, while the effects appeared in a total dose of 60 to 70 mg and further effects were obtained after about 1 week of the termination of the therapy. Leukopenia occurred in 6 of 14 cases. The side effects of THP-adriamycin were milder than those of ADM.

Free-flow isotachophoresis under micro-gravity.

When free flow electrophoresis (FFE) is applied to the separation and purification process under microgravity, higher separability is expected than that under usual 1 G environment, because sedimentation of samples and heat convection can be neglected. Although FFE under micro-gravity is a very important technique for efficient separation of macromolecules because of the above merits, FFE unit (FFEU) should be more sophisticated to give stable operation, high separability, high through-put and high separation efficiency. In this study, fundamental research to achieve FFE with such high performance was done by using a commercial FFEU. The research contains (1) the quantitative study on the separation mode of FFE and separation efficiency, (2) the feasibility analysis of free-flow bidirectional isotachophoresis (FFBITP) which enables separation of cations anions simultaneously by isotachophoresis (ITP) and zone electrophoresis (ZE), (3) the optimization of hardware factors such as thickness of the separation chamber which may cause deterioration of separation due to heat convection, and (4) the analysis of hardware factors depending on the long-time operation. The ultimate goal of this study is to obtain fundamental data for the development of FFE for the international space station. In this paper, the experimental results obtained in a reduced-gravity aircraft was mainly.

[The effects of fresh gas flow rate on the work of breathing during semi-closed circuit anesthesia].

To evaluate the effects of fresh gas flow rate on the work of breathing during semi-closed circuit anesthesia, we measured respiratory frequency, tidal volume, inspiratory work of breathing, peak inspiratory and expiratory flow rate and fluctuations in airway pressure during breathing with fresh gas flow of 6 l.min-1, 3 l.min-1 and 1.5 l.min-1. There was no significant difference among three flow rates in terms of tidal volume, respiratory frequency, inspiratory work of breathing, peak inspiratory flow rate and fluctuation in airway pressure at inspiration. On the other hand, in the expiratory phase, peak expiratory flow rate was lower and fluctuation in airway pressure at expiration was higher at 6 l.min-1 than those at 3 l.min-1 and 1.5 l.min-1, which may be indicative of increased expiratory work of breathing. The present study shows that low fresh gas flow rate does not increase the inspiratory work of breathing and high fresh gas flow rate may increase the expiratory work of breathing, implying that lowering the fresh gas flow is profitable for the work of breathing during semi-closed circuit anesthesia.

[THP-adriamycin in head and neck tumor].

Primary effects and side-effects of a new anthracycline antineoplastic agent, THP-adriamycin (THP-ADM), were examined in 20 patients with malignant head and neck tumors. According to a classification by tumor sites, there were 6 cases of oropharynx, 6 of nose and sinuses, 3 of tongue, 2 of floor of mouth, 2 of neck and 1 of external auditory meatus. According to the tissue classification, there were 11 cases of squamous cell ca., 6 cases of malignant lymphoma, each one case of anaplastic ca., carcino sarcoma and adeno cystic ca. There were 16 previously untreated cases and 4 pretreated cases. Twelve cases received THP-ADM by intraarterial injection and 8 cases by the same dose. 10 to 20 mg/body 3 times a week, in a total of 40 to 100 mg. 2 CR, 5 PR, 3 MR and 4 NC in 14 cases with carcinoma, and 2 CR, 3 PR and 1 MR in 6 cases with malignant lymphoma were obtained. Among 12 cases receiving intraarterial injection, 3 CR and 5 PR were obtained. The decrease of WBC counts below 3000/mm3 after THP-ADM administration was observed in 9 cases. Side effect of THP-ADM appeared to be less severe than that of Adriamycin.

[Clinical results of FAR (5-FU, vitamin A, and radiation) therapy in head and neck cancer].

Combination therapy of 5-FU, vitamin A and radiation (FAR therapy) was given to 50 patients with squamous cell carcinoma of the head and neck. The primary effect was observed in 22 patients who received FAR therapy as radical therapy. Among them, only 4 patients had recurrence or metastasis, and death was none. FAR therapy was given to 4 patients as pre-operative therapy and to 2 patients as post-operative therapy. However, the number of cases was not enough for us to draw any conclusion. In the group of 22 patients consisting of patients with recurrence or metastasis and patients who refused operation or were unresectable, no primary effect was observed only in 4 patients. Among a total of 50 patients, only 4 patients did not show a primary response to FAR therapy, 19 had relapse, 19 died (6 cases of them died from other causes), and 31 are still alive. There were no patients having severe side effects. From these results, FAR therapy seemed to be worth trying in treatment of head and neck cancer.

[Phase II study of UFT for head and neck cancer].

A Phase II study of UFT for head and neck cancer was conducted in 10 institutions. UFT is a mixture of Futraful and uracil. Eighty-four patients entered this trial, of which 60 were evaluable. UFT was administered orally at a daily dose of 600 mg/day. Eight patients achieved complete response and 10 achieved partial response with an over-all response rate of 30.0 %. Evaluating response according to by histology, the response rate was 30.9% for cases of squamous cell carcinoma. Complete response was observed in one case of undifferentiated carcinoma. Response rate according to primary site was 33 to 40% for the nose & paranasal sinuses, mesopharynx, hypopharynx and larynx. The response rate was 28.9% for the group of patients treated previously, and 33.3% for the group previously untreated. The mean time for 50% or more regression of the tumor was 4.3 weeks. Toxic effects appeared in 40.3% of 67 evaluable cases as anorexia, nausea, vomiting, stomatitis, diarrhea etc. In one case of maxillary carcinoma, severe bone marrow suppression was observed. We concluded that UFT therapy was markedly effective for head and neck cancer.

[Moderate dose methotrexate with citrovorum factor rescue therapy in the treatment of head and neck cancer].

A clinical trial of moderate dose methotrexate (MTX)-CF rescue was conducted in 12 institutions. Thirty-seven patients with head and neck carcinoma entered this trial, of which 32 were evaluable. MTX was administered 350 mg/m2 (500 mg/body) by i.v. drip over 6 hours. Three hours after completion of MTX infusion, CF rescue was started. There was no complete response in 32 patients. Nine patients showed partial response with the response rate of 28%. The response rates were 21% for the group of patients treated previously, and 75% for the group untreated previously. MTX concentration in plasma was determined at 6, 24, 48 and 72 hours after the initiation of MTX infusion, and the assay results revealed a safe range. GI disturbances were seen at the rates of 11 to 38%. Bone marrow suppression was mild and no renal toxicity was observed. We concluded that the moderate dose MTX-CF rescue therapy was useful for head and neck carcinoma. As a next step, we are planning to conduct a clinical trial of high-dose MTX.