Prevention of chemotherapy toxicity by agents that neutralize or degrade cell-free chromatin.

BACKGROUND: Toxicity associated with chemotherapy is a major therapeutic challenge and is caused by chemotherapy-induced DNA damage and inflammation. We have recently reported that cell-free chromatin (cfCh) fragments released from dying cells can readily enter into healthy cells of the body to integrate into their genomes and induce DNA double-strand breaks, apoptosis and inflammation in them. We hypothesized that much of the toxicity of chemotherapy might be due to release of large quantities of cfCh from dying cells that could trigger an exaggerated DNA damage, apoptotic and inflammatory response in healthy cells over and above that caused by the drugs themselves. METHODS: We tested this hypothesis by administering cfCh neutralizing/degrading agents namely, anti-histone antibody complexed nanoparticles, DNase I and a novel DNA degrading agent-Resveratrol-Cu concurrently with five different chemotherapeutic agents to examine if chemotherapy-induced toxicity could be minimized. RESULTS: We observed (i) significant reduction in chemotherapy-induced surge of cfCh in blood; (ii) significant reduction in chemotherapy-induced surge of inflammatory cytokines CRP, IL-6, IFNγ and TNFα in blood; (iii) abolition of chemotherapy-induced tissue DNA damage (γH2AX), apoptosis (active caspase-3) and inflammation (NFκB and IL-6) in multiple organs and peripheral blood mononuclear cells; (iv) prevention of prolonged neutropenia following a single injection of adriamycin and (v) significant reduction in death following a lethal dose of adriamycin. CONCLUSION: Our results suggest that toxicity of chemotherapy is caused to a large extent by cfCh released from dying cells and can be prevented by concurrent treatment with cfCh neutralizing/degrading agents.

Development of a new wastewater treatment process for resource recovery of carotenoids.

A new wastewater treatment process that involves coagulation, ozonation, and microalgae cultivation has been developed. Here, two challenges are discussed. The first was minimizing phosphorus removal during coagulation in order to maximize algal production. The second was to optimize microalgae cultivation; algal species that grow rapidly and produce valuable products are ideal for selection. Haematococcus pluvialis, which produces the carotenoid astaxanthin, was used. Growth rate, nutrient removal ability, and astaxanthin production of H. pluvialis in coagulated wastewater were investigated. After coagulation with chitosan, the turbidity and suspended solids decreased by 89% ± 8% and 73% ± 16%, respectively. The nitrogen and phosphorus contents of the supernatant remained at 86% ± 6% and 67% ± 24%, respectively. These results indicate that coagulation with chitosan can remove turbidity and SS while preserving nutrients. H. pluvialis grew well in the supernatant of coagulated wastewater. The astaxanthin yield from coagulated wastewater in which microalgae were cultured was 3.26 mg/L, and total phosphorus and nitrogen contents decreased 99% ± 1% and 90% ± 8% (Days 31­35), respectively [corrected].

Seasonal changes in the performance of a catch crop for mitigating diffuse agricultural pollution.

An in situ technology for mitigating diffuse agricultural pollution using catch crops was developed for simultaneously preventing nitrate groundwater pollution, reducing nitrous oxide (N2O) gas emissions, and removing salts from the topsoil. Seasonal changes in the performance of a catch crop were investigated using lysimeters in a full-scale greenhouse experiment with 50 d cultivation of dent corn. Catch crop cultivation significantly reduced the leached mineral nitrogen by 89-91% in summer, 87-89% in spring, and 61-82% in winter, and it also significantly reduced the N2O emission by 68-84% in summer. The amounts of nitrogen uptake by the catch crop were remarkably higher than those of leached nitrogen and N2O emission in each season. Catch crop cultivation is a promising technology for mitigating diffuse agricultural pollution.

Nutrient recovery from biomass cultivated as catch crop for removing accumulated fertilizer in farm soil.

As a result of long-term continuous use of fertilizers in farm land, a large amount of nutrients accumulate in the soil, increasing the risk of eutrophication or nitrate pollution of groundwater. For rehabilitating the farm soil and recovering nutrients such as nitrogen, phosphorus and potassium, a new system has been developed by our research group. This paper discusses the methodology of extracting nutrients from biomass in order to recover phosphorus and other nutrients in crystal form. Around 80% or higher extraction rates were achieved for phosphorus and potassium by soaking the powdered tissue in distilled water or 1% NaOH solution for 24 h. The extracted phosphorus and potassium act as a potential resource for recycled fertilizer or other industrial materials.

Operation of a new sewage treatment process with technologies of excess sludge reduction and phosphorus recovery.

This paper shows the potential application of a new sewage treatment process with technologies of excess sludge reduction and phosphorus recovery. The process incorporated ozonation for excess sludge reduction and crystallisation process for phosphorus recovery to a conventional anaerobic/oxic (A/O) phosphorus removal process. A lab-scale continuous operation experiment was conducted with the ratio of sludge flow rate to ozonation tank of 1.1% of sewage inflow under 30 to 40 mgO3/gSS of ozone consumption and with sludge wasting ratio of 0.34% (one-fifth of a conventional A/O process). Throughout the operational experiment, a 60% reduction of excess sludge production was achieved in the new process. A biomass concentration of 2300 mg/L was maintained, and the accumulation of inactive biomass was not observed. The new process was estimated to give a phosphorus recovery degree of more than 70% as an advantage of excess sludge reduction. The slight increase in effluent COD was observed, but the process performance was maintained at a satisfactory level. These facts demonstrate an effectiveness of the new process for excess sludge reduction as well as for phosphorus recovery.

Advanced sewage treatment process with excess sludge reduction and phosphorus recovery.

An advanced sewage treatment process has been developed, in which excess sludge reduction by ozonation and phosphorus recovery by crystallization process are incorporated to a conventional anaerobic/oxic (A/O) phosphorus removal process. The mathematical model was developed to describe the mass balance principal at a steady state of this process. Sludge ozonation experiments were carried out to investigate solubilization characteristics of sludge and change in microbial activity by using sludge cultured with feed of synthetic sewage under A/O process. Phosphorus was solubilized by ozonation as well as organics, and acid-hydrolyzable phosphorus (AHP) was the most part of solubilized phosphorus for phosphorus accumulating organisms (PAOs) containing sludge. At solubilization of 30%, around 70% of sludge was inactivated by ozonation. The results based on these studies indicated that the proposed process configuration has potential to reduce the excess sludge production as well as to recover phosphorus in usable forms. The system performance results show that this system is practical, in which 30% of solubilization degree was achieved by ozonation. In this study, 30% of solubilization was achieved at 30 mgO(3)/gSS of ozone consumption.

[Comparison of in vitro antimicrobial activities of ofloxacin, levofloxacin, ciprofloxacin, and sparfloxacin against various mycobacteria].

In vitro antimicrobial activities of ofloxacin (OFLX), levofloxacin (LVFX), ciprofloxacin (CPFX), and sparfloxacin (SPFX) were compared against various mycobacteria using the agar dilution method with 7H11 medium, and the following results were obtained. (1) These four new quinolones showed excellent antimicrobial activities against M. tuberculosis, M. kansasii, and M. fortuitum. (2) SPFX was most active against slowly growing mycobacteria. The activity against M. tuberculosis was in the order of SPFX > CPFX > LVFX > OFLX. The activity against M. kansasii was in the order of SPFX > LVFX > OFLX > or = CPFX. (3) On the other hand, CPFX was most active against rapidly growing mycobacteria. The activity against M. fortuitum was in the order of CPFX > SPFX > LVFX > OFLX. Considering the in vitro antimicrobial activities and the pharmacokinetics of these four drugs, they could achieve favorable clinical outcomes for all the patients with pulmonary infection due to M. tuberculosis or M. fortuitum and some of the patients with pulmonary infection due to M. kansasii or M. chelonae.

Water quality characteristics of forest rivers around Lake Biwa.

Forest river surveys were carried out at upper streams of several rivers in the Lake Biwa watershed to understand the water quality characteristics of the rivers, and to find out their relationships with forest features such as geographical, geological and vegetational data. The results showed: (1) Forests have some purification functions for nitrogen and organic matter, but become sources for most of ionic species. (2) Main mineral species in forest rivers are Ca2+, Mg2+ and Na+, HCO3-, CO3(2-), Cl-, SO4(2-) and SiO2. (3) Loading from forests was 0.4-7 kg/km2/d for TN and 0.01-0.3 kg/km2/d for TP. (4) River quality reflects the properties of each forest, and is unique to the place, especially in ionic species such as Ca2+ and Cl-. (5) A cluster analysis successfully categorized ionic components into several groups.

[In vitro antimycobacterial activities of a new quinolone, balofloxacin].

Balofloxacin (BLFX), a newly developed fluoroquinolone, was studied for its in vitro antimycobacterial activity by the agar dilution method with 7H11 agar medium. The MIC90s were as follows: 0.39 microgram/ml for M. tuberculosis, > 50 micrograms/ml for M. avium, > 50 micrograms/ml for M. intracellulare, 0.39 microgram/ml for M. kansasii, 0.39 microgram/ml for M. fortuitum, > 50 micrograms/ml for M. abscessus, and 50 micrograms/ml for M. chelonae. The antimycobacterial activity of BLFX was comparable or slightly inferior to that of levofloxacin (LVFX). Considering the present findings and pharmacokinetics of BFLX, it appears that BFLX may achieve favorable outcome in the treatment of patients with infection due to M. tuberculosis, M. kansasii, or M. fortuitum similar to that of ofloxacin or LVFX.

[Clinical evaluation of new quinolones as antituberculosis drugs].

Compared with the recent rapid advances in the diagnosis of tuberculosis, advances in the treatment of tuberculosis have been quite slow. For example, as long as six months are required for initial treatment, even with addition of pyrazinamide in the first two months to isoniazid, rifampicin, and streptomycin or ethambutol. Moreover, it is not always easy to treat patients who cannot receive standard agents including isoniazid and rifampicin due to adverse effects of these drugs or drug resistance. For these reasons, the development of new agents with potent antituberculosis activities and fewer adverse effects is urgently desired. However, at present, few new antituberculosis agents are being developed, and new quinolones are considered the most promising new antituberculosis agents due to their lack of cross-resistance to previously existing antituberculosis agents, their excellent in vitro and in vivo antituberculous activities, and good pharmacokinetics. We therefore reviewed experimental studies and clinical reports useful for evaluation of potential clinical use of new quinolones as antituberculosis drugs. Our conclusions are summarized below: 1) Of nine new quinolones on the market, ofloxacin, ciprofloxacin, aparfloxacin and levofloxacin have excellent in vitro and in vivo antituberculous activities without cross-resistance to previously existing antituberculosis agents. 2) Ofloxacin appears to be useful clinically for intractable multidrug-resistant tuberculosis. The incidence and severity of adverse effects of ofloxacin were very low on longterm administration. 3) Ofloxacin resistance emerged from two to four months after initiation of administration of ofloxacin, and ofloxacin exhibited cross-resistance to other new quinolones. 4) The usefulness of new quinolones for initial treatment of tuberculosis is unclear. 5) New quinolones should be used to treat patients with drug-resistant tuberculosis and those patients for whom adverse effects have limited the use of standard agents. However, monotherapy with new quinolones is not recommended, due to the significant risk of emergence of quinolone resistance. 6) Not only ofloxacin, ciprofloxacin, sparfloxacin and levofloxacin, which are on the market, but also gatifloxacin, CS-940, Du-6859a and other newly developed new quinolones are candidates for new antituberculosis agents. However, careful study not only of antituberculous activities and pharmacokinetic but also drug interactions and chronic cumulative toxicities due to long-term administration are needed prior to clinical application of these drugs.

[Nontuberculous mycobacteriosis; the present status and in the future. 3--(1). The view of development of new drugs against nontuberculous mycobacterial infections].

It is obvious that the number of patients with pulmonary nontuberculous mycobacterial infections is increasing gradually in Japan. Of these infections, M. avium complex (MAC) is the most common cause, and is known to be resistant to many antimicrobial drugs. At present, no standard regimen which is able to control MAC infections completely is established. For these reasons, the development of new drugs with strong antimycobacterial activity which are not cross-resistant to conventional antimycobacterial drugs is urgently desired. Thus, we studied in vitro activities of various drugs which are expected to be a new promising drug against nontuberculous mycobacterial infections, and reviewed clinical impact of these drugs. 1) New quinolones New quinolones including ofloxacin, ciprofloxacin, levofloxacin and sparfloxacin (SPFX), are considered to be active against M. tuberculosis, M. kansasii, M. fortuitum, but are inactive against MAC, M. chelonae, M. abscessus, M. scrofulaceum. Both AM-1155 and Du-6859a, newer quinolones, seemed to be comparable to or more active than SPFX which is considered to be most active now. 2) New macrolides Clarithromycin (CAM) has in vitro activities against various nontuberculous mycobacteria including MAC, and also has proven to have clinical potential not only for disseminated MAC infections in AIDS but also for pulmonary MAC infections. Therefore, CAM seems to be a candidate for one of the key drugs in the treatment of MAC infections. 3)Rifamycins Rifabutin (RBT) and rifapentine exhibited more potent in vitro and in vivo antimycobacterial activities than rifampicin. RBT has already demonstrated clinical effect against intractable tuberculosis and MAC infections. Thus, RBT is recommended for the prophylaxis of M. tuberculosis and MAC infections in AIDS patients in US. KRM-1648 displayed much more potent in vitro and in vivo activities than rifampicin against both M. tuberculosis and MAC. It is needed an effort to confirm its therapeutic efficacies. Now clinical phase study is going on in US. 4) Phenazines Clofazimine (CFZ), an effective antileprosy drug, is known to be active in vitro against various mycobacteria including MAC, and often used as a component of combination chemotherapy for disseminated MAC infections in AIDS patients in US. Recently, CFZ new analogs have been developed, and it is necessary to evaluate its activities against nontuberculous mycobacteria.

[Study on purple pigmentation in five cases with purple urine bag syndrome].

We reported five patients with purple urine bag syndrome (PUBS). Four patients had indicanuria, and, in three of them, purple pigmentation was reproduced by inoculating urinary isolates, in the broth with indoxyl sulfate. Klebsiella pneumoniae, Pseudomonas aeruginosa and Enterococcus avium were considered to produce the purple pigment in three patients. However, attempts to reproduce the purple pigment failed in two patients, and one of them did not have indicanuria. These results suggest that indicanuria is not necessarily required for the development of PUBS.

[Therapeutic potential of sparfloxacin for preventing mycobacterial infections].

We studied the therapeutic potential of utilizing sparfloxacin (SPFX), a newly developed quinolone, to prevent various mycobacterial infections. The in vitro activity of SPFX as a preventive agent for various mycobacteria was determined using the actual count method on Ogawa egg medium. The minimal inhibitory concentrations (MICs) of SPFX were as follows: ofloxacin-sensitive M. tuberculosis, 0.16-0.32 microgram/ml; ofloxacin-resistant M. tuberculosis, 0.63-2.5 micrograms/ml; M. avium; 0.63-10 micrograms/ml (MICs were equal or less than 1.25 micrograms/ml in seven out of 11 strains); M. intracellulare, 2.5-10 micrograms/ml (MICs were equal or more than 10 micrograms/ml in 17 out of 23 strains); M. kansasii, < or = 0.08-0.16 microgram/ml; M. fortuitum, < or = 0.08 microgram/ml; M. chelonae subsp. abscessus, > 10 micrograms/ml; M. chelonae subsp. chelonae, 0.63 microgram/ml; M. scrofulaceum, < or = 0.08 microgram/ml; M. nonchromogenicum, 1.25 micrograms/ml; M. xenopi, < or = 0.08 microgram/ml; M. gordonae, < or = 0.08 microgram/ml. The average serum concentrations of SPFX during the period of multiple oral administration (200 mg once a day) were 0.35 +/- 0.16 microgram/ml before administration, 0.67 +/- 0.32 microgram/ml after one hour, 1.13 +/- 0.21 microgram/ml after two hours, 1.27 +/- 0.32 microgram/ml after four hours and 1.31 +/- 0.34 micrograms/ml after six hours. These results indicate that SPFX has a strong therapeutic potential to prevent infections due to M. tuberculosis, M. kansasii, M. fortuitum, M. chelonae subsp. chelonae, M. scrofulaceum, M. xenopi and M. gordonae. Moreover, it may be expected to be a promising agent against infections due to ofloxacin-resistant M. tuberculosis, M. avium and M. nonchromogenicum.

[Flora in the anterior nares of patients in the neurologic ward: the influence of nasogastric intubation and mechanical ventilation].

MRSA has been isolated frequently from patients in the neurologic ward of Minami-Okayama Hospital. We examined the flora in the anterior nares of 37 patients in the neurologic ward and 36 patients in the medical ward. Existence of dysphagia and nasogastric intubation is considered to make a marked difference between nasal flora of neurologic patients and that of medical patients. Patients with nasogastric intubation had higher incidences of colonization with S. aureus, and gram-negative rods. All patients colonized with MRSA had received nasal tube-feeding. Nasogastric intubation is considered to be a significant risk factor for MRSA colonization. The study in patients with nasogastric intubation demonstrated that patients with mechanical ventilation had a significantly higher incidence of colonization with gram-negative rods, such as P. aeruginosa, and a lower incidence of colonization with S. aureus. A incidence of MRSA colonization in patients with mechanical ventilation was significantly low. Mechanical ventilation is not necessarily considered to be a risk factor for MRSA colonization.