Induction of thymidine phosphorylase expression and enhancement of efficacy of capecitabine or 5'-deoxy-5-fluorouridine by cyclophosphamide in mammary tumor models.

Thymidine phosphorylase (dThdPase) is an essential enzyme for the activation of the oral cytostatic drugs capecitabine (N(4)-pentyloxycarbonyl-5'-deoxy-5-fluorocytidine, Xeloda(trade mark)) and its intermediate metabolite doxifluridine [5'-deoxy-5-fluorouridine (5'-dFUrd, Furtulon((R)))] to 5-fluorouracil (5-FUra) in tumors. In a previous study, we found that several cytostatics were able to up-regulate tumor levels of dThdPase in a human colon cancer xenograft model. In the present study, we confirmed that the administration of cytostatics used for breast cancer treatment, such as taxanes and cyclophosphamide (CPA), up-regulated the tumor level of dThdPase in mammary tumor models as well. Because the dThdPase up-regulation was observed even when CPA was given orally, we investigated further the usefulness of combination therapy with the 2 oral drugs, 5'-dFUrd/capecitabine and CPA in mammary tumor models. Daily oral administration of CPA up-regulated human dThdPase levels in the tumor tissue of mice bearing a human mammary tumor xenograft, MX-1, whereas in the small intestine and liver, it did not affect levels of pyrimidine nucleoside phosphorylases (PyNPase) including dThdPase and uridine phosphorylase. The preferential up-regulation of PyNPase activity in the tumor by CPA administration was also confirmed in mice bearing a syngeneic murine mammary adenocarcinoma, A755. In both models, combination therapy of 5'-dFUrd/capecitabine with CPA showed synergistic antitumor activity, without significant potentiation of toxicity. In contrast, treatment with CPA and either 5-FUra or UFT (a mixture of tegafur and uracil) in combination showed only additive activity. Our results suggest that CPA and capecitabine/5'-dFUrd, both available for oral administration, would be good partners, and that clinical trials with this drug combination against breast cancer are warranted.

On the prevalence of Asiatic cholera in Okinawa, 1879 and "Ryukyu-kiko" Written by Hironobu Tsuchiya.

The greatest prevalence of Asiatic cholera since the adoption of modern statistics was in 1879. Especially in Okinawa Prefecture, its morbidity rate was the largest in Japan in that year. Dr. Hironobu Tsuchiya, who was appointed as an official of the Department of Inner Affairs, wrote "Ryukyu-kiko" as a private memorandum. He wrote more about the situation of administrative confusion than he did about the condition of Asiatic cholera. This paper also mentions the brief sketch of Tsuchiya's life and the description of the manufacturing methods, effectiveness and use of chemical drugs which are contained in his "Sinyaku-shoko".

Antitumor activities of a novel fluoropyrimidine, N4-pentyloxycarbonyl-5'-deoxy-5-fluorocytidine (capecitabine).

Capecitabine (N4-pentyloxycarbonyl-5'-deoxy-5-fluorocytidine) is a novel fluoropyrimidine carbamate that was synthesized for the purpose of finding antitumor drugs with improved safety and efficacy profiles compared with those of 5-fluorouracil (5-FUra) and doxifluridine (5'-deoxy-5-fluorouridine, 5'-dFUrd). The present study compared the antitumor activities of the compound with those of other fluoropyrimidines in 12 human cancer xenograft models and their antimetastatic activities in murine tumor models. The antitumor efficacy of capecitabine was greater than those of 5'-dFUrd, UFT (a mixture of tegafur and uracil) and 5-FUra. Capecitabine was also much safer, particularly much less toxic to the intestinal tract, than the other compounds, indicating higher therapeutic indices. The therapeutic indices of capecitabine, 5'-dFUrd and 5-FUra were >40, >20 and 2.0 against the human CXF280 colon cancer xenograft, the most sensitive line to the fluoropyrimidines so far tested, and 5.1, 1.5, and <1.5 against the human HCT116 colon cancer xenograft with ordinary sensitivity, respectively. In addition, capecitabine, as well as 5'-dFUrd, selectively suppressed the spontaneous metastasis of mouse Lewis lung carcinoma in mice at extremely low doses, 32-64 fold lower than their minimum effective dose (MED) against the primary tumor growth. Capecitabine was even more antimetastatic than 5'-dFUrd. These results indicate that capecitabine has high therapeutic potential.

Basic studies on hydroxy apatite cement: I. Setting reaction.

Self-setting cements, alpha D-Cement and alpha DT-Cement, were prepared. They consisted of only the calcium phosphates alpha-TCP, TTCP and DCPA. These cements reacted and hardened in a moist environment at 37 degrees C. The powder X-ray diffraction patterns were taken to examine the conversion of their reactions as a function of time. The cements reacted and produced hydroxyapatite. The optimum powder/liquid ratio of alpha D-Cement was 2.0 and that of alpha DT-Cement was 1.8. The initial setting time of alpha D-Cement was 87.5 m and that of alpha DT-Cement was 107.5 m. The component and the product of these cements are calcium phosphates which are the putative minerals in teeth and bones. Therefore, these cements are useful for oral surgery as bone-filling materials.

Tumor selective delivery of 5-fluorouracil by capecitabine, a new oral fluoropyrimidine carbamate, in human cancer xenografts.

Capecitabine (N4-pentyloxycarbonyl-5'-deoxy-5-fluorocytidine) is a novel fluoropyrimidine carbamate that is converted to 5-fluorouracil (5-FUra) by three enzymes located in the liver and tumors; the final step is the conversion of 5'-deoxy-5-fluorouridine (5'-dFUrd) to 5-FUra by thymidine phosphorylase in tumors. The present study compared the efficacy of capecitabine and 5-FUra at their maximum tolerated doses in CXF280, HCT116, COLO205, and WiDr human colon cancer xenograft models, and measured subsequent 5-FUra and 5'-dFUrd levels in tumors and in the plasma and muscle. Capecitabine was effective in the first three models, whereas 5-FUra was effective only in CXF280, which is a cell line highly susceptible to fluoropyrimidines. In the three susceptible models, 5-FUra AUCs in tumors after capecitabine administration were 210 to 303 nmol x hr/g, whereas those after 5-FUra administration were 8.54 to 13.1 nmol x hr/g. In addition, capecitabine gave higher levels of 5-FUra AUC in tumors than in plasma (114- to 209-fold higher) and muscle (21.6-fold higher), whereas 5-FUra was not selectively distributed to tumors. In the refractory model, WiDr, 5-FUra AUC in tumors after capecitabine administration was only 62.8 nmol x hr/g, although the level of the intermediate metabolite 5'-dFUrd was high (AUC: 695 nmol x hr/g). The ratio of 5-FUra/5'-dFUrd levels in the WiDr tumors was 0.09, which was 23.8-fold lower than that in the HCT116 tumors. The mechanism of resistance would be the inefficient conversion of 5'-dFUrd to 5-FUra by thymidine phosphorylase in tumors. Thus, capecitabine might show its high efficacy as a result of delivering high levels of 5-FUra selectively to the tumors.

Induction of thymidine phosphorylase activity and enhancement of capecitabine efficacy by taxol/taxotere in human cancer xenografts.

Thymidine phosphorylase (dThdPase) is an essential enzyme for the activation of the cytostatics capecitabine (N(4)-pentyloxycarbonyl-5'-deoxy-5-fluorocytidine) and its intermediate metabolite [5'-deoxy-5-fluorouridine (5'-dFUrd)] to 5-fluorouracil in tumors. We have tried to identify the best partners of capecitabine in combination therapy, such as dThdPase up-regulators, which may enhance the efficacy of this compound. Among various cytostatics studied with the WiDr human colon cancer xenograft model, Taxol, Taxotere, and mitomycin C greatly increased levels of human dThdPase in tumors, and cyclophosphamide slightly increased the enzyme level. These cytostatics simultaneously increased the levels of human tumor necrosis factor alpha (TNFalpha), which is an up-regulator of dThdPase. In cultures of the WiDr cells, however, Taxol did not up-regulate TNFalpha to a detectable level and only slightly enhanced levels of dThdPase. These results suggest that Taxol might indirectly elevate TNFalpha in tumor cells, which in turn up-regulated dThdPase in the tumor cells in the WiDr cancer xenograft. In the combination therapy, the efficacy of Taxol and Taxotere with either capecitabine or 5'-dFUrd was more than just additive. In contrast, Taxol and either 5-fluorouracil or UFT (a mixture of tegafur and uracil) in combination showed only additive activity. Taxol and Taxotere might enhance the efficacy of capecitabine and 5'-dFUrd, probably by modulating dThdPase activity in tumor tissues.

Positive correlation between the efficacy of capecitabine and doxifluridine and the ratio of thymidine phosphorylase to dihydropyrimidine dehydrogenase activities in tumors in human cancer xenografts.

Capecitabine (N4-pentyloxycarbonyl-5'-deoxy-5-fluorocytidine) is a new fluoropyrimidine carbamate, which is converted to 5-fluorouracil (5-FUra) selectively in tumors through the intermediate metabolite 5'-deoxy-5-fluorouridine (5'-dFUrd, doxifluridine). 5'-dFUrd is metabolized to 5-FUra by thymidine phosphorylase (dThdPase) located in high levels in various types of solid tumors from patients, whereas 5-FUra generated is catabolized to dihydrofluorouracil by dihydropyrimidine dehydrogenase (DPD). The present study investigated whether the efficacy of capecitabine and its intermediate metabolite 5'-dFUrd correlates with levels of these enzymes in various human cancer xenograft models. Capecitabine and 5'-dFUrd were highly effective and inhibited tumor growth by more than 50% in 18 of 24 xenograft lines (75%) and 15 of 24 xenograft lines (63%), respectively, whereas 5-FUra and a mixture of tegafur and uracil were effective only in 1 of 24 (4.2%) and 5 of 24 (21%), respectively. The efficacy of capecitabine correlated with dThdPase activity. However, capecitabine was effective even in tumors with lower levels of dThdPase if DPD levels were also lower. In contrast, it was not as effective even in tumors with sufficient levels of dThdPase if DPD levels were very high. The efficacy of capecitabine consequently correlated very well with and depended on the ratio of these two enzymes in tumors. These results indicate that capecitabine might exert its efficacy through 5-FUra generated in tumor tissues but not through that generated in normal organs. On the other hand, there was no correlation between the efficacy of a mixture of tegafur and uracil and these enzyme activities in tumors. The efficacy of capecitabine would be optimized by selecting patients who have tumors with a high ratio of dThdPase to DPD activities.

Immunohistochemical staining of human teeth, and production of monoclonal antibodies against cementum.

Influence of various forms of fixation and decalcification on immunohistochemical staining of paraffin-embedded human teeth and surrounding tissues was initially examined using commercially available antibodies against vimentin (V), type I collagen (C) and cytokeratin (K). Secondly, monoclonal antibody (MoAb) was produced against both bovine and human cementum and immunohistochemical screening was subsequently undertaken to test reactivity with different scheme of fixation and decalcification. The combination of neutral buffered paraformaldehyde fixation, Morse solution and unmasking procedure yielded both optimal morphology and immunoreactivity. The application of this method into production of MoAb against human or bovine cementum generated a variety of MoAbs reactive with both human teeth and surrounding tissues. The percentage of hybridoma supernatant reactive with sections of human teeth was 14.0-22.4% and was both higher and much improved compared to results of previous soft tissue studies. Finally, the MoAbs, BC 1 and 2 (isolated following the use of bovine cementum as immunogen) and HC 1 (human cementum immunogen) recognized specific bands of various molecular weight. The three MoAbs showed strong resistance against periodate oxidation and borohydrate reduction and were stable following treatment with proteolytic enzymes. All were immunoreactive with components of cementum.

[Dr. Torafumi Okuyama, naval medical officer and the author of the dictionaries of medical terms].

Dr. Genryo Torafumi Okuyama, the second son of Dr. Genchu Okuyama of the Kaminoyama clan, was born on Dec. 4th, 1847. His elder brother Dr. Toraakira Okuyama was promoted to Dai Ikan (Senior Captain), the highest rank of medical officer in the Japanese Navy, and rendered distinguished services in the establishment of the naval medical systematization in the early Meiji era. Dr. Trafumi Okuyama, who was appointed as medical officer of the Yokohama army Hospital and transferred to Daibyoin in Edo, was engaged in medical treatment of injured soldiers during the Boshin-war in 1868. He went to Kagoshima with William Willis and as one of the founders of the Kagoshima Medical school, gave students education there. He resigned his naval position in 1874, when he was Dai Gun I (Senior Leutenant) and died at the age of 41 in April 16th 1887. Dr. Torafumi Okuyama compiled A medical vocabulary in English and Japanese ("Igo Ruizyu") and Deutsch-Japanisches Hand-Wörterbuch für Medizin ("Dokuwa Igaku Ziten) and published "Koen Hikki", the translation of the lectures by Dr. Edwin Wheeler.

[Discord between the Department of Defense and Toko on the foundation of the Bureau of the Army Doctors].

The relation between the Department of Defense and Toko was not intimate before the Bureau of Army Doctors, his relation to Toko was also not smooth. and then he had to employ the army doctors from the organization other than Toko. I pointed out that the transfer of the higher army doctors to the Bureau of the Army Doctors was done not just after its foundation, as it was said before, but in 1874, after the normalization of the relationship of the two.

[Dr. Toraakira Okuyama, a higher medical officer of the Japanese navy in Meiji era].

Toraakira Okuyama was born in Nagasaki in 1840. His father, Genchu Okuyama, was the Dutch learning doctor of the Kaminoyama clan and one of the founders of the Otamagaike Vaccination Center. During Tokugawa period T. Okkuyama, who had a title "Gensei", was appointed as medical officer in the infantry regiments in 1863 and then was promoted to vice-director. He was appointed as doctor of the national hospital (Daibyoin) and then as assistant professor in the new epoch of Meiji was transferred to become a naval medical officer in 1871. He was advanced to Dai-ikan, the highest rank of medical officer in the Japanese navy. He worked for the establishment of naval medical systematization and gave students education at the naval medical school (Kaigun-Guni-Ryo-Gakusha) with William Anderson and Edwin Wheeler. He resigned his post in 1876 and died in 1926, until which time he continued the life of a practitioner.

Effects of cementum-impregnated gelatine membrane on early healing following periodontal flap surgery.

The purpose of this study was to investigate the initial healing response of surgically flapped periodontal tissues in the presence of gelatine membrane compounded with particles of cementum. Four monkeys with no periodontal disease were used in this experiment. Full thickness flaps were raised and recession type defects were created on the buccal side of the maxillary lateral incisors and second premolars. Exposed root surfaces were thoroughly curetted, and composite cementum-impregnated membranes placed directly onto planed root surfaces. Flaps were then sutured back to the original position. Animals were sacrificed at 2, 4, 7 and 10 post-surgical days, and block specimens including the tooth, gingiva and bone were subsequently processed for light and electron microscopy. The resultant analysis revealed that gelatine membranes were partially resorbed at 2 days post-surgery and completely resorbed by 10 postoperative days. In the early stages of gelatine resorption, most liberated cementum particles accumulated on planed dentin surface but some became demineralized within the surgical wound. Cementoblast-like cells with well-developed rough endoplasmic reticulum appeared on the root surface 7 days following surgery. Newly synthesized collagen fibrils aligned parallel to the root surface were seen at 10 post-surgical days. The results suggest that the newly developed composite membrane enhances the formation of new periodontal attachment.

Neutrophil activating activity of tonsillar cells from patients with tonsillitis.

The previous paper demonstrated that tonsillar cells cultured in vitro in the presence or absence of a streptococcal preparation, OK-432, produce factors that activate various neutrophil functions. In the present study, examination was made of the factor productivity of tonsillar cells from patients with chronic tonsillitis of varying severity, and palmoplantar pustulosis (PPP). Tonsillar cells from patients with severe tonsillitis and PPP incubated with culture medium alone produced a much greater amount of active factors compared with those from patients with mild tonsillitis. When tonsillar cells were incubated in the presence of OK-432, augmentation in factor production by the addition of OK-432 was less in former than latter cases, suggesting that factor production from tonsils correlates with the course of inflammation in this organ.

Effects of supernatant from tonsillar cell culture on neutrophil functions.

The palatine tonsil is the most important element of Waldeyer's ring as a defense mechanism against various microorganisms. The neutrophils are the initial active participants in bacterial infection and inflammation, but their interaction with other cellular participants is poorly understood. To clarify this point, the effects of culture supernatant from human tonsillar cells on the peripheral neutrophil function were investigated. Supernatants from tonsillar cells were incubated in the presence or absence of streptococcal preparation (OK-432) enhanced chemiluminescence, adherence, phagocytosis, chemotaxis, and superoxide production of human peripheral neutrophils. The results suggest that tonsils play an important role in the regulation of neutrophil function.

Thermal coefficients of paste-paste type pulp capping cements.

Thermal coefficients of four kinds of commercially available paste-paste type pulp capping cements were examined. Control reference samples were made of dentin. A thermal coefficient analyzer was used, heating specimens for a few nanoseconds by a xenon flash bulb and measuring thermal changes by using a thermocouple. Thermal coefficients were examined by this non-steady state method. Thermal conductivities of all cements were almost the same or lower than that of dentin. Therefore, when each cement was heated, the penetrating energy was almost the same or lower than that of dentin. The thickness of the cements was converted into that of the dentin by using the obtained thermal conductivity. The 1-mm thickness of the examined cements were equal to between 0.97-mm and 2.10-mm thicknesses of lost dentin. The use of a pulp capping cement provided better pulp protection from thermal stimuli than did the same thickness of dentin.

Effects of hyperlipaemic serum on interleukin-2 (IL-2) production, IL-2 receptor expression, and T cell proliferation induced by IL-2 in cynomolgus monkeys.

The effect of hyperlipaemic serum on mitogen-induced T lymphocyte proliferation was investigated with cynomolgus monkeys. The mitogen-induced blastogenesis was remarkably inhibited when either hyperlipaemic or normal monkey lymphocytes were incubated with hyperlipaemic sera. Hyperlipaemic serum also inhibited ConA-induced interleukin 2 (IL-2) production as well as IL-2 receptor (IL-2R) expression of normal monkey lymphocytes. On the other hand, it showed slight inhibition of T-cell proliferation induced by adding recombinant human IL-2 to IL-2R-positive normal monkey lymphocytes. These results indicate that hyperlipaemic serum inhibited an early stage of T-cell autocrine activation pathway including IL-2 production and IL-2R expression.

Setting reactions and compressive strengths of calcium phosphate cements.

Setting reactions and compressive strengths of a self-hardening calcium phosphate cement (CPC) were investigated. The CPC consists of tetracalcium phosphate (TTCP) and anhydrous dicalcium phosphate (DCPA). The cement specimens were prepared by mixing 0.7 g of the powder (TTCP 72.9 wt% + DCPA 27.1 wt%) with 0.175 mL of the liquid (25 mmol/L H3PO4 and 1.32 mmol/L sodium fluoride). The specimens were removed from the molds at pre-determined time intervals after being mixed, and their compressive strengths were measured. Immediately afterward, the fractured specimens were rapidly frozen in ethanol (-80 degrees C), lyophilized, and examined by powder x-ray diffraction and scanning electron microscopy (SEM). The results showed that (1) hydroxyapatite was the only reaction product; (2) the reaction was nearly completed within four h, during which both the reaction product and compressive strength increased linearly with time, resulting in a strong correlation between the two; and (3) fully set CPC consisted primarily of small rod-like crystals and some platy crystals.