Search

Background/aim: The objective of this study was to investigate the significance of the blood levels of free amino acids (AAs) in infertile men., Patients and Methods: Ninety-three men who underwent examinations for infertility were included. The concentrations of 20 AAs were measured and compared in four groups (normospermia, obstructive azoospermia, oligozoospermia, non-obstructive azoospermia) based on semen analysis and clinical parameters., Results: When the 93 men were divided into normospermia, obstructive azoospermia, oligozoospermia, and non-obstructive azoospermia groups, no significant differences were found in the concentrations of the 20 AAs between them. We then compared 49 men diagnosed with normozoospermia or oligozoospermia according to the median sperm motility and morphology abnormalities rates (30% and 20%, respectively). Men with low motility rates had significantly lower levels of tryptophan and alanine, and men with high abnormal morphology rates had significantly lower levels of aspartate and glutamate., Conclusion: AAs are probably involved in the pathogenesis of male infertility, particularly oligozoospermia., (Copyright© 2021, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Background/aim: We investigated whether mastication affects microglia, whose activity is thought to be associated with cognition and brain tumor progression., Materials and Methods: We kept mice by feeding either a hard or soft diet for 2, 4 or 8 months. After each period, we removed the whole brains and isolated microglia. The total RNA extracted from each brain's microglia was subjected to DNA microarray analysis., Results: Many genes were found to be significantly differentially expressed between hard- and soft-diet-fed mice in each group of the same feeding period. The expression of several genes involved in the regulation of actin cytoskeleton was down-regulated in the soft-diet-fed mice., Conclusion: Mastication may affect microglia's roles in cognition as well as their neuroimmune activity through their activity of patrolling the brain., (Copyright© 2020, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Background: Oral mucositis (OM) induced by cancer chemotherapy has a high incidence and serious symptoms, which often force chemotherapy to be stopped. GGsTop is a newly-discovered gamma-glutamyl transpeptidase (GGT) inhibitor. Previous research suggested that inhibition of GGT suppressed reactive oxygen species and induced the production of collagen and elastin. We hypothesized that GGsTop could safely treat OM., Materials and Methods: A mouse model of OM was treated with GGsTop and ulcer area, weight, and white blood cell count were determined. The treatment effect was also evaluated by hematoxylin-eosin and collagen staining., Results: The therapeutic effect of GGsTop was better than that of an existing drug and may be safely used in combination with chemotherapy. Furthermore, GGsTop promoted collagen production in oral mucosa., Conclusion: GGsTop treated OM quickly and safely. GGsTop is highly valuable for use as a treatment for OM., (Copyright© 2019, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Background: Oral mucositis (OM), one of the side-effects induced by chemotherapy, has 40% incidence and the incidence rate increases to approximately 100% in combination with radiotherapy. We describe OM in ICR mice induced using 5-fluorouracil (5-FU) and 20% acetic acid., Materials and Methods: We optimized the dose of 5-FU and 20% acetic acid and validated the efficacy of standard therapies for OM., Results: All mice developed OM after administration of 5-FU and 20% acetic acid. Application of Kenalog® reduced maximum ulcer area and the duration of spontaneous recovery in a dose-dependent manner., Conclusion: We succeeded in developing a mouse model of OM induced by cancer chemotherapy. New drugs for OM induced by anticancer drugs can be evaluated simply by monitoring the WBC count in this mouse model. This model is expected to contribute to development of new drugs and elucidation of the mechanisms of ameliorating stomatitis as a side-effect of anticancer drugs., (Copyright© 2018, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Background/aim: Clinically used disinfectants are often irritating and cause skin problems. Ozone water is unique among disinfectants. It does not damage skin cells and readily decomposes to oxygen without generating harmful residues. On the other hand, it rapidly loses its sanitizing activity. Recently developed nano-bubble ozone water (NBOW) can keep its sanitizing activity much longer. This study aimed to examine the microbicidal effects of NBOW after long-term storage., Materials and Methods: The concentration of ozone in NBOW was examined by measuring the NBOW redox potential. Microbicidal activity was evaluated by colony formation assays, after incubating bacteria with NBOW for set time periods., Results: NBOW lost its microbicidal activity after 1 year of storage at 4°C. Stocked frozen, NBOW retained appreciable microbicidal activity after 1 year of storage. Mycobacterium smegmatis, one of the most disinfectant-resistant bacteria, was killed within 15 min. NBOW was resistant to freeze-thawing., Conclusion: NBOW that had been stored frozen possessed sufficient microbicidal activity to kill bacteria even after 1 year of storage. Moreover, it was shown that NBOW is freeze-thaw resistant. NBOW possesses desirable features rendering it an attractive alternative disinfectant., (Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Background: Polymer drug delivery systems designed to reduce systemic side-effects are clinically important. Polyphosphoesters are biodegradable polymers with versatile structure that could afford reactive sites or polar functions for drug immobilization., Materials and Methods: The drug-polyphosphester systems were characterized by nuclear magnetic resonance and infrared spectroscopy, differential scanning calorimetry and dynamic light scattering. In vitro and in vivo experiments were performed to assess the biological activity of the immobilized drug., Results: Two water-soluble polyphosphoester-based delivery systems of paclitaxel were synthesized. The conjugate with paclitaxel covalently bonded to the polymer, had attenuated activity in vitro. The second system comprised of macromolecular aggregates incorporating the drug via hydrogen bonding. The physical complex achieved a certain level of antitumor activity in vivo and no decrease of body weight - evidence for reduction of the systemic toxic effect associated with paclitaxel treatment., Conclusion: The physical complex was found to be a promising carrier for delivery of toxic anticancer agents, e.g. paclitaxel., (Copyright© 2016 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.)

Background: Group-specific component (Gc)-globulin-derived macrophage-activating factor (GcMAF) generated by a cascade of catalytic reactions with deglycosidase enzymes exerts antitumor activity. We hypothesized that degalactosyl Gc-globulin (DG3), a precursor of GcMAF, also plays a role in recovery from cancer as well as GcMAF due to progression of deglycosylation by generally resident sialidases and mannosidases., Materials and Methods: We prepared the subtypes of DG3, such as 1f1f and 1s1s and its 22 homodimers, by using vitamin D3-binding Sepharose CL-6B and examined their antitumor activity in mice bearing Lewis lung carcinoma cells, by counting the number of nodules formed in their lungs., Results: Antitumor activity of DG3 was observed regardless of its subtype, being equivalent to that of GcMAF. The injection route of DG3 affected its antitumor activity, with subcutaneous and intramuscular administration being more favorable than the intraperitoneal or intravenous route. In order to obtain significant antitumor activity, more than 160 ng/kg of DG3 were required., Conclusion: DG3 proved to be promising as an antitumor agent, similarly to GcMAF.

Background: The 1f1f subtype of the group-specific component (Gc) protein is converted into Gc protein-derived macrophage-activating factor (GcMAF) by enzymatic processing with β-galactosidase and sialidase. We previously demonstrated that preGc(1f1f)MAF, a full Gc(1f1f) protein otherwise lacking a galactosyl moiety, can be converted to GcMAF by treatment with mouse peritoneal fluid. Here, we investigated the effects of the β-galactosidase-treated 1s1s and 22 subtypes of Gc protein (preGc(1s1s)MAF and preGc₂₂MAF) on the phagocytic activation of mouse peritoneal macrophages., Results: We demonstrated the presence of Gal-GalNAc disaccharide sugar structures in the Gc(1s1s) protein by western blotting using peanut agglutinin and Helix pomatia agglutinin lectin. We also found that preGc(1s1s)MAF and preGc₂₂MAF significantly enhanced the phagocytic activity of mouse peritoneal macrophages in the presence and absence of mouse peritoneal fluid., Conclusion: We demonstrate that preGc(1s1s)MAF and preGc₂₂MAF proteins can be used as effective macrophage activators.

Background: The 1f1f subtype of the Gc protein (Gc(1f1f) protein) was converted into Gc-derived macrophage-activating factor (GcMAF) by enzymatic processing in the presence of β-galactosidase of an activated B-cell and sialidase of a T-cell. We hypothesized that preGc(1f1f)MAF, the only Gc(1f1f) protein lacking galactose, can be converted to GcMAF in vivo because sialic acid is cleaved by residual sialidase. Hence, we investigated the effect of preGc(1f1f)MAF on the phagocytic activation of mouse peritoneal macrophages., Results: We examined the sugar moiety of preGc(1f1f)MAF with a Western blot using peanut agglutinin (PNA) and Helix pomatia agglutinin (HPA) lectin. We also found that preGc(1f1f)MAF significantly enhanced phagocytic activity in mouse peritoneal macrophages but only in the presence of the mouse peritoneal fluid; the level of phagocytic activity was the same as that observed for GcMAF., Conclusion: PreGc(1f1f)MAF can be used as an effective macrophage activator in vivo.

Aim: It is possible that macrophages may be effective for cancer treatment because once activated, lung macrophages have enhanced contact with lung tumor cells and have a cytotoxic effect. In this paper, we report that nitric oxide (NO) produced by lung macrophages activated with lipopolysaccharide (LPS) suppressed cell growth of human lung adenocarcinoma cells., Materials and Methods: A549, a lung adenocarcinoma cell line, was cultured with NR8383, a rat alveolar macrophage cell line, in the presence and absence of LPS. The effect of LPS on the growth rate of A549 cells was examined as a function of NO production under cell-to-cell contact conditions., Results: NR8383 cells showed potent cytostatic and cytocidal effects on A549 cells when both cells were co-cultured in the presence of LPS. These effects were mainly due to the production of NO, but another possible mechanism, such as cell-to-cell contact, may also be involved., Conclusion: Activation of alveolar macrophages by LPS suppresses the growth of lung carcinoma cells via NO production under cell-to-cell contact conditions.

Background: In lung tumor biotherapy, local macrophages such as alveolar macrophages and tumor-associated macrophages (TAMs) which normally exist in contact with tumor cells are thought to be hopeful target. It is advantageous to clarify the potential for and mechanism by which lung tumor cells are killed by the neighboring macrophages in order to establish new lung cancer therapy using a drug delivery system to lung tissue., Materials and Methods: A549, a human lung adenocarcinoma cell line, and Lewis lung carcinoma LLC1, a mouse lung cancer cell line, were co-cultured with NR8383, a rat alveolar macrophage cell line, and AMJ2-C11, a mouse alveolar macrophage cell line, at a ratio of 1:10 and 1:5, respectively. Macrophages were activated with lipopolysaccharide (LPS) and cytotoxicity toward tumor cells was evaluated by a dye-uptake method and (3)H-thymidine release assay, respectively. Nitric oxide (NO) production was estimated by Griess assay, and tumor necrosis factor (TNF)-α and interleukin (IL)-1β were measured by ELISA., Results: Significant macrophage cell aggregation and cytotoxicity against A549 cells and LLC1 cells was observed with NR8383 cells and AMJ2-C11 cells in the presence of LPS. A high concentration of NO and TNF-α were detected in the supernatant of co-culture medium with LPS. Inhibition of cell-to-cell contact restored A549 cell growth., Conclusion: The LPS-activated alveolar macrophages demonstrated an increased cell-to-cell contact with lung tumor cells and inducing cytotoxicity with production of NO and cytokines TNF-α and IL-1β. These results suggest that moderate activation of local macrophages in lung (alveolar macrophages and TAMs) is thought to be a hopeful means of establishing new immunotherapy for lung cancer.

Background: Micro- and nanospheres have been used as carrier materials for effective delivery of drugs into macrophages via endocytosis including phagocytosis for the treatment of cancer and infections., Materials and Methods: To analyze the role of lipid rafts in the endocytic uptake of microspheres by macrophages, we studied the incorporation of polystyrene latex microspheres (PSL MS) by treating macrophages with reagents that affect the physicochemical and biochemical functions of their lipid rafts, such as the cholesterol depletor methyl-β-cyclodextrin (MβCD) and the inhibitor of scavenger receptors polyinosinic acid (poly I). For this study, we used 3-μm fluorescent polystyrene latex microspheres (F-PSL MS) and J774 murine macrophage., Results: We found that the endocytosis of J774 cells was remarkably reduced in a concentration-dependent manner by treatment of the cells with MβCD and that endocytosis was suppressed by approximately 50% by treatment with 100 μg/ml poly I. These results suggest that scavenger receptors are associated with phagocytosis of F-PSL MS and that their functions are regulated by lipid rafts., Conclusion: PSL MS are phagocytosed via a raft-dependent pathway.

Aim: We performed a retrospective analysis of a calcium hydroxide-containing calcium agent (TACHIKAWA DENKAI CALCIUM™: E-Ca) based on data of bone mineral density obtained by dual-energy X-ray absorptiometry (DXA) to clarify the relationship between bone mineral density and E-Ca intake on an empty stomach in those who regularly use E-Ca., Results: We found the percentage of volunteers with their age-matched (AM) values of above 100% to be 89%, and also a moderate positive correlation between AM values and the intake period of E-Ca., Conclusion: Our findings demonstrate that AM values can be used as an effective indicator assessing osteogenesis by regularly administrated calcium agents which exert their effects after long-term use.

Background: Macrophage cells differentiate into killer macrophages, named M1 macrophages, that effectively eliminate cancer cells by generating cytokines. We examined the feasibility of a lung cancer therapy using lipopolysaccharide (LPS), which triggers this differentiation. It is expected that the delivery via inhalation of LPS directly into the lungs, where alveolar macrophages reside abundantly, would be effective at minimizing the possible toxic effect of LPS., Materials and Methods: We determined the effects of intratracheal insufflation of LPS on (i) the exudation of lactate dehydrogenase, (ii) generation of tumor necrosis factor-α and interleukin-12, and (iii) tumor metastases in Lewis lung carcinoma-bearing C57BL/6 mice., Results: Pulmonary insufflation of LPS resulted in a consistent accumulation of tumor necrosis factor-α, and transient increase in interleukin-12 without significant release of lactate dehydrogenase from the lung cells. In addition, a significant antitumor effect of LPS was observed; and this antitumor effect was potentiated by combination of LPS with the antitumor agent cyclophosphamide., Conclusion: Pulmonary inhalation of LPS combined with a chemotherapeutic agent is a promising approach to lung cancer therapy.

Background: Over the last decade, tumor-specific antigens have been discovered, but so far it has not been possible to use them as part of an effective acquired immunotherapy. This failure may be due to the fact that the expression of the MHC class 1 is low and in lung cancer cells is heterogeneous. Therefore, it may be advantageous to develop techniques that activate the antitumor mechanism of the innate immune system. An experimental model was developed for testing lung cancer therapies that are based on the stimulation of macrophages, which then activate innate immunity., Materials and Methods: A549, a human lung adenocarcinoma cell line, was co-cultured with a rat macrophage cell line (NR8383), or a human macrophage cell line (THP 1) at the ratios of 1:1 or 1:5. The experiments were performed with lipopolysaccharide (LPS) or in its absence. The cytotoxicity rate to A549 cells was estimated over time using a dye-uptake method and the amount of lactate dehydrogenase released was measured. The amount of nitric oxide (NO) induced in the medium was assayed, because it may be a candidate as a useful cytotoxic factor., Results: High cytotoxicity was observed to A549 cells when co-cultured with NR8383 cells in the presence of LPS. This effect was not observed in the absence of LPS. Similar results, although to a lesser extent, were observed when A549 cells were co-cultured with THP-1 cells. A high concentration of NO was measured in the co-culture medium of A549 cells and NR8383 cells when LPS was present., Conclusion: The induction of cell death in lung cancer cells occurred after contact with macrophages that had been activated by LPS. The NO that was produced by macrophages in response to LPS was responsible for some of this effect.

Background: Mycobacterium tuberculosis infection affects one-third of the world's population and causes the death of three million people each year. To clarify details of M. tuberculosis survival strategies, it is important to establish a suitable in vitro model that mimics a chronic infection in alveolar macrophages by M. tuberculosis. For this reason, we established a new in vitro model using a rat alveolar macrophage cell line, NR8383., Materials and Methods: Basic characteristics, including phagocytotic ability and production of nitrogen oxide and tumor necrosis factor in response to several stimuli, of NR8383 cells were compared with those of primary alveolar macrophages. The course after phagocytosis of live or killed M. bovis bacilli Calmette-Guerin (BCG) was examined over 21 days using NR8383 cells as the host., Results: The characteristics that have been examined to date were nearly the same for both primary alveolar macrophage and NR8383 cells. Live BCG phagocytosed by NR8383 cells had successfully begun to grow in the cells within 7 days, while killed BCG were almost completely destroyed by 21 days., Conclusion: BCG-infected NR8383 cells are potentially a suitable in vitro model that mimics a chronic infection with M tuberculosis.

Appropriate and rational modulation of innate immunity may enhance the therapeutic efficacy of emerging immune therapies for treating cancer. One of the crucial cells of innate immunity is the macrophage. The purpose of this article was to review those issues that suggest ways of exploiting macrophage local functions in immune therapy, and to discuss the suitability of low molecular-weight lipopolysaccharides as potent modulators of macrophage functions for immune therapy of cancer.