Distribution of gallium-67 in normal and hypotransferrinemic tumor-bearing mice

The mechanism by which 67Ga accumulates in tumors is controversial. The most popular theory is that 67Ga binds to transferrin and gains access to cells by the transferrin receptor. However, substantial evidence suggests that uptake of 67Ga may not be universally mediated by transferrin in tumors. To determine whether transferrin is required for uptake of 67Ga in vivo, we compared the uptake of 67Ga by two types of implanted tumors and by normal tissues in normal and severely hypotransferrinemic strains of Balb/C mice. One type of tumor was strongly gallium-avid in normal mice; the other was not. Uptake of 67Ga by normal soft tissues was markedly less in hypotransferrinemic than in normal mice. Uptake of 67Ga by bone was equivalent in the two types of mice. For the more gallium-avid tumor, uptake of 67Ga was similar and the ratio of tumor-to-background activity was substantially higher in the hypotransferrinemic than in the normal mice. For the less gallium-avid tumor, uptake was significantly less in hypotransferrinemic than in normal mice. These data suggest that uptake of 67Ga by bone and by some tumors may be a transferrin-independent process.     

Plasma hormone levels in normal and lead-treated pregnant mice

Dietary lead (0.5%) was given to mice which, after mating, exhibited a vaginal plug. Estradiol, progesterone and prostaglandins E and F 2 alpha were determined in the plasma by radioimmuno assay at different times thereafter. The increase in estradiol and decrease in prostaglandins prior implantation are not greatly altered by lead treatment, whereas the subsequent increase in progesterone and later in estradiol is abolished. It is concluded that the lower number of pregnancies seen in lead-treated mothers is due to a maternal hormonal imbalance caused by lead.     

Coenzyme Q10 and adriamycin toxicity in mice

Pretreatment for four days with coenzyme Q10 (CoQ10) significantly lowered the acute toxicity in female C3H/HeNCrlBR mice given moderately lethal (15.0 and 20.0 mg/kg) i.p. doses of adriamycin as well as in male ICR/Hla mice given 12.5 mg/kg i.p. adriamycin. In both strains of mice, CoQ10 pretreatment did not protect the mice at higher i.p. adriamycin dose levels. When adriamycin was administered by the clinically-used i.v. route, CoQ10 pretreatment did not reduce acute toxicity at moderately lethal doses in either strain. At higher i.v. adriamycin dose levels, CoQ10 pretreatment significantly enhanced acute toxicity. CoQ10 pretreatment did not alter the antitumor effectiveness of adriamycin (i.p. or i.v.) against the Dunn osteosarcoma.     

Continuous infusion of 5-fluorouracil plus low-dose cisplatin in tumor-bearing mice

We previously reported that the topical application of ascorbic acid 2-O-alpha-glucoside (AA-2G) suppressed the cutaneous inflammation by ultraviolet irradiation in human and guinea pigs (Miyai et al., Nishinihon J. Dermatol., 58, 439-443 (1996)). In this paper, the effect of AA-2G on the lethal damage induced by ultraviolet B (UVB) was studied using a human keratinocyte cell line, SCC, established from squamous cell carcinoma. The photoprotective effect of AA-2G on cytotoxicity of UVB in SCC cells was dose dependent (0.125-1 mM) and more effective than that of ascorbic acid (AsA) at 1 mM. This protection was completely abolished in the presence of an alpha-glucosidase inhibitor, castanospermine, indicating that release of AsA from this derivative was essential for reduction of the actinic injury. AA-2G significantly suppressed cytotoxicities of hydrogen peroxide and superoxide anion produced by xanthine and xanthine oxidase. AA-2G exhibited a preventive effect against the cytotoxicity produced by tert-butylhydroperoxide, an inducer of lipid peroxidation, in the presence of alpha-tocopherol, but not in the absence of alpha-tocopherol. Cytotoxicity of UVB was also effectively reduced by the combination of AA-2G and alpha-tocopherol. In addition, AA-2G reduced UVB-promoted formation of lipid peroxide and accumulation of lipofuscin, which is known to be a complex of cellular proteins and metabolites of lipid peroxide. These data suggest that AA-2G prevents the acute inflammation induced by UVB irradiation partly through scavenging reactive oxygen species and potentiating the antioxidative activity of alpha-tocopherol.     

Emergence of higher levels of invasive and metastatic properties in the drug resistant cancer cell lines after the repeated administration of cisplatin in tumor-bearing mice

To establish a more suitable model for reflecting biological aggressiveness in clinically recurrent cancers after chemotherapy, we made the in-vitro-established cisplatin-resistant cell lines, by exposing the parental tumor cell lines to cisplatin in a culture system, and also the in-vivo-established cisplatin-resistant cell lines by repeated cisplatin administration to parental tumor-bearing mice. Although both cell lines similarly demonstrated a clinically relevant low level of drug resistance (from 1.5 to 2.9 times more resistance to cisplatin than their parental cell lines), only the in-vivo-established cisplatin-resistant cell lines showed significantly enhanced metastatic properties with a 2.1- to 3.4-fold increase in the number of lung metastatic nodules. These enhanced metastatic properties were caused by tumor invasiveness in combination with various levels of enhancement of cell attachment, proteolytic enzyme activity and cell motility. We concluded that anticancer drugs such as cisplatin could promote tumor progression only in the drug-resistant cell lines established in vivo. As a result, these cell lines are considered to be a more faithful and useful model for expressing biological aggressiveness in clinically recurrent cancers after chemotherapy than the conventional drug-resistant cell lines established in vitro.     

Regulation on beta-END in tumor-bearing mice by moxibustion on Guanyuan point

The experimental results demonstrated that moxibustion on Guanyuan point could promote the hyperplasia of the pituitary and the adrenal gland which showed atrophy in control group. Also, the moxibustion treatment stimulated the secretion of beta-END from the pituitary and the adrenal gland, increased the level of serum beta-END significantly and kept the high level for quite a long time, that was advantageous for beta-END to carry out the immunomodulation. The data further suggested that moxibustion treatment do not cause the instant release but probably constant release of the beta-END.     

Pepstatin, an ascites retardant of L1210 tumor-bearing mice

The effect of pepstatin on the kinetics of ascitic fluid accumulation in L1210 tumor-bearing mice (DBA/2) was observed. Following inoculation of 1.5x10(6) tumor cells, untreated mice reached a peak of fluid accumulation on day 6 and remained at this level until death on day 9. A "lag" phase of 4 days occurred before fluid accumulation was seen. Pepstatin administered SC in a single dose of 80 mg/kg during the lag phase, significantly retarded fluid accumulation as compared to untreated animals. Pepstatin administered following fluid accumulation was much less effective. We concluded that pepstatin prevents fluid accumulation rather than acts as a diuretic agent. The term "ascites retardant" is suggested for the pharmacologic actions of pepstatin, since it prevents fluid accumulation without diminishing the cell count.     

Time dependent cytotoxicity of adriamycin and daunomycin in primary cultures of normal and neoplastic mammary glands

In the ongoing NCDS, 44 children of the 46 identified at 7 years as having a serious unilateral hearing loss were followed up at II years. Half of them had recovered normal bilateral hearing, the remainder still had serious deafness in one ear. Although at 7 years the 46 children as a group had shown backwardness in oral ability, speech and reading, at II years both the "recovered" and the "persistent" were similar to their age peers in scholastic attainment. Despite their original apparent difficulty it was encouraging to find that several children in both subgroups were noted as possessing outstanding academic ability. It is concluded that with prompt follow-up by an alerted school doctor, children with unilateral deafness at age 7 years are likely to progress satisfactorily in later childhood.     

Heart rate variability in insomniacs and matched normal sleepers

BACKGROUND: Heat shock proteins have been associated with the mutant form of the tumor suppressor gene, TP53, and with resistance to cancer chemotherapy. METHODS: Archival tissues from 50 patients with head and neck squamous cell carcinoma who received primary surgical resection were examined for p53, HSP27, and HSP70 by immunohistochemistry and correlated with tumor stage, grade, and 5-year survival (alive or deceased). RESULTS: Both heat shock proteins were strongly expressed in normal mucosa and in small (T1 and T2) tumors. Thirty (60%) of tumors were positive for p53, 43 (86%) for HSP27, and 34 (68%) for HSP70, with no association between p53 and heat shock protein expression. Twenty-five patients were alive (4 with disease), and 25 patients were deceased (9 from other causes). p53 Protein overexpression correlated with low-grade tumors. Only primary tumor site (i.e., oral cavity > larynx > oropharynx/base of tongue) and N stage were significantly associated with survival. CONCLUSIONS: Heat shock proteins are expressed in normal upper respiratory tract squamous mucosa, and their role in carcinoma remains unclear. None of the markers, p53, HSP27, or HSP70, demonstrated prognostic significance for 5-year survival. We confirm the recognized association of cervical lymph node metastases with decreased survival.     

Reduction by coenzyme Q10 of the acute toxicity of adriamycin in mice

Pretreatment for four days with coenzyme Q10 (COQ10) reduced the acute toxicity in mice treated with adriamycin. In two sequential protocols, adriamycin allowed only 36 and 42% survival, respectively. Pretreatment with COQ10 allowed 80 and 86% survival, respectively. The differences are significant, p less than 0.05. The mechanism for this reduction in the acute toxicity may be based upon the prevention by the supplementary COQ10 of the inhibition caused by adriamycin to COQ10-dependent enzymes in cardiac and and other tissues. The prospect of diminishing the toxicity of adriamycin in cancer patients remains promising and important.     

白血病肿瘤疫苗联合1-甲基色氨酸免疫治疗荷瘤小鼠疗效观察

目的 探讨白血病肿瘤疫苗(简称瘤苗)主动免疫治疗及联合吲哚2,3双加氧酶(IDO)的抑制剂1-甲基色氨酸(1-MT),在白血病荷瘤小鼠治疗中的作用.方法 采用FBL-3细胞皮下注射建立荷瘤白血病小鼠模型;实验分为5组:正常对照组、PBS对照组、环磷酰胺(CTX)化疗组、单用瘤苗治疗组和瘤苗联合1-MT治疗组;观察各组小鼠的一般状况、肿瘤缓解率、肿瘤大小、转移情况及生存期.结果 PBS对照组小鼠活动迟缓,体质量(含瘤结节质量)比其余各组均高;单用瘤苗组和瘤苗联合1-MT组小鼠活动、进食正常,体质量与正常小鼠筹异不大;化疗组体质量明显减轻,出现脱毛、弓背、活动减少等,差异有统计学意义(F=57.71,P=000);单用瘤苗组和瘤苗联合1-MT组治疗相关死亡率明显低于化疗组(0,0,40%).瘤苗联合1-MT组完全缓解率与单用瘤苗组(61.1%、70.0%)比较,差异无统计学意义(χ2=0.221,P>0.05),但瘤苗联合1-MT组的复发率低于单用瘤苗组(0,36.36%);复发小鼠再应用1-MT,能明显抑制瘤结节的生长.单用瘤苗组和瘤苗联合1-MT组小鼠中位存活期明显高于化疗组和PBS对照组(χ2=52.13,P<0.01).各组小鼠整体瘤结节的变化比较差异有统计学意义(F=89.966,P=0.000).结论 白血病瘤苗在动物实验具有肯定的疗效,能明显抑制肿瘤的生长,延长小鼠生存时间,且副作用小.免疫治疗联合1-MT对白血病进行治疗,可以显著减少肿瘤的复发率;而免疫治疗有效后复发时应用1-MT,可以显著抑制肿瘤的生长.     

Realization of the therapeutic potential of CTLA-4 blockade in low-dose chemotherapy-treated tumor-bearing mice

CTLA-4 blockade has been shown by other investigators [D. R. Leach, et al., Science (Washington DC), 271: 1734-1736, 1996; and Y-F. Yang, et al., Cancer Res., 57: 4036-4041, 1997] to retard tumor growth in selected tumor systems. Here, we show that CTLA-4 blockade alone was ineffective in retarding tumor growth in the murine MOPC-315 tumor system. Yet, CTLA-4 blockade offered significant therapeutic benefits to MOPC-315 tumor bearers when combined with a subtherapeutic dose of the chemotherapeutic agent melphalan, which was previously shown (L. Gorelik, et al., Cancer Immunol. Immunother., 39: 117-126, 1994) to shift the cytokine profile in the tumor bearers toward type-1 cytokines. In addition, we show here that anti-CTLA-4 monoclonal antibody enhanced antitumor cytotoxicity when the anti-CTLA-4 monoclonal antibody was added to stimulation cultures of spleen cells from low-dose melphalan-treated MOPC-315 tumor-bearing mice but not from untreated tumor-bearing mice. These results suggest that the therapeutic benefits of CTLA-4 blockade depend on the ability of drugs such as melphalan to promote an immunogenic environment by altering the cytokine profile of tumor-specific T cells.     

Melatonin decreases bone marrow and lymphatic toxicity of adriamycin in mice bearing TLX5 lymphoma

When CBA male mice bearing TLX5 lymphoma were treated in the evening with a single i.v. dose of adriamycin (20-40 mg/Kg), the administration of a single pharmacological dose of melatonin (10 mg/kg s.c.) 1 hr earlier reduced the acute mortality from 10/24 to 2/24. The increase in survival time caused by adriamycin over drug untreated controls was not reduced by melatonin. The administration of melatonin alone did not cause any antitumor or evident toxic effect. Melatonin also attenuated the reduction caused by adriamycin in the number of bone marrow GM-CFU, and of CD3+, CD4+ and CD8+ splenic T-lymphocyte subsets. Reduced and total glutathione levels were decreased in the bone marrow and in the liver cells of the animals treated with adriamycin, and were significantly restored by melatonin. Moreover, lipid peroxidation by adriamycin was reduced by melatonin, as indicated by malondialdehyde measurement in the liver of the treated animals. These data indicate that the protective effects of melatonin against the host toxicity of the prooxidant antitumor drug, adriamycin, might be attributed at least partially to its antioxidant properties. These findings appear of interest in relation to the physiological rhythmic levels of endogenous melatonin and to the chronotoxicology of anthracyclines.     

Immunoregulation by interleukin-12 in MB49.1 tumor-bearing mice: cellular and cytokine-mediated effector mechanisms

Administration of recombinant murine interleukin (rmIL)-12 to MB49.1 tumor-bearing mice results in dose-dependent regression of the primary tumor and the generation of protective antitumor immunity in the majority of animals. rmIL-12 administration is associated with a marked increase in lymph node cellularity that is predominantly due to the expansion of B220+ B cells as well as CD8+ T cells. Stimulation of lymph node cells from rmIL-12-treated, but not control tumor-bearing mice, with MB49.1 tumor cells in vitro was shown to enhance the secretion of interferon (IFN)-gamma. The magnitude of this in vitro response was dependent on the dose of rmIL-12 administered in vivo and mirrored the change in circulating serum IFN-gamma. Furthermore, at the height of the in vitro response to tumor stimulation, the addition of a neutralizing antibody to murine IL-12 suppressed IFN-gamma production, indicating a role for endogenous IL-12 in this antigen-specific cytokine response. Although studies in SCID mice confirmed that an appropriate T cell response was required for rmIL-12-mediated antitumor activity, in immunocompetent animals early tumor regression was not accompanied by cellular infiltration of the tumor. In contrast, a profound increase in tumor-associated inducible nitric oxide synthase (iNOS) was observed in mice receiving rmIL-12 which preceded T cell infiltration of the tumor which could be detected during the second week of IL-12 treatment. Direct tumor killing through the cytotoxic actions of NO via the iNOS pathway may serve as a way of generating tumor antigen which enables the host to mount a subsequent T cell response against the tumor.     

Maternal plasma D-dimer levels in normal and complicated pregnancies

OBJECTIVE: To evaluate D-dimer as a marker for fibrinolysis in normal and complicated pregnancies using an enzyme-linked immunosorbent assay (ELISA) technique. METHODS: Four groups of pregnant women were enrolled: 17 normal women followed longitudinally from 28-40 weeks' gestation, 14 patients with preterm labor at 28-34 weeks, 17 patients with preeclampsia at term (37-40 weeks), and 14 patients with abruptio placentae (32-40 weeks). We assayed peripheral venous blood samples from each patient for D-dimer levels using a commercial ELISA kit. D-dimer values were calculated by regression analysis using internal standards and controls for each assay. Data were compared using Student t test or analysis of variance with repeated measures. RESULTS: D-dimer values increased slightly with increasing gestational age. Patients with preterm labor, preeclampsia, and abruptio placentae had mean D-dimer values significantly greater than those of controls (P < .003). D-dimer values of the abruption group were approximately twice those of the control group (3393 +/- 2086 versus 1750 +/- 839 ng/dL). CONCLUSION: An increase in fibrinolysis may be associated with the pregnancy complications studied, as reflected by alterations in maternal plasma D-dimer levels.     

Nitrite and nitrate levels in cerebrospinal fluid of normal subjects

In order to evaluate the involvement of nitric oxide in neurologic disorders it is important to generate controlled values of its metabolites nitrite and nitrate in human cerebrospinal fluid (CSF). Samples of CSF obtained from 14 patients without neurologic diseases were analysed for nitrite and nitrate concentration by reverse phase chromatography with ultraviolet (UV) detection. For comparison, the levels of nitrite in the same samples were also measured by reverse phase chromatography coupled with electrochemical detection and those of nitrate by ion chromatography coupled with UV detection. A good correlation was found for the concentration values of both ions obtained with the two procedures. Then, 10.41 +/- 0.47 ng/ml of nitrite and 2.92 +/- 0.37 ng/ml of nitrate could be regarded as reliable values in control subjects. No correlation between age and levels of nitrite and nitrate was observed.     

Distribution of adriamycin in mice under conditions of local hyperthermia which improve systemic drug therapy

Local tumor hyperthermia (LTH) (43 degrees C for 60 minutes) enhanced the effectiveness of Adriamycin (ADR) (10 mg/kg iv) administered systemically against the 16C mouse mammary adenocarcinoma. LTH was produced by the local application of 2450 MHz microwaves to sc implanted tumors. During LTH the intratumor temperature uniformity was +/- 0.5 degrees C and the rectal temperatures were 36 degrees C--38 degrees C. The simultaneous application of LTH and ADR resulted in tumor growth delays which were greater than those expected from the addition of individual treatments. Enhanced effectiveness of these treatments did not occur if the two treatments were separated by 4 hours. Cures were observed only in the treatment groups that received drug and heat simultaneously. Measurements of the tissue distribution of ADR and its metabolites using 14C-ADR indicated that the uptake of radioactivity in the heated tumors was not increased by LTH. A greater percentage of the radioactivity in heated tumors was recovered as ADR metabolites and correspondingly less radioactivity was recovered as unchanged ADR.