Application Of Cyclophosphamide Research Articles

The effects of berberine and curcumin on cardiac, lipid profile and fibrosis markers in cyclophosphamide-induced cardiac damage: The role of the TRPM2 channel.

Cyclophosphamide (CYP) is widely used to treat various types of cancer. In addition to the therapeutic properties of this drug, unfortunately, its side effects are still not fully understood. This study investigated the protective effect of curcumin (CURC) and berberine (BER) on CYP-induced cardiac damage. Thirty-six male rats were equally divided into the control, dimethyl sulfoxide (DMSO), CYP, CYP + CURC, CYP + BER and CYP + BER + CURC groups. Troponin-I, Creatine kinase-myocardial band (CK-MB), total cholesterol, triglyceride levels in serum samples, and reactive oxygen species (ROS), poly(ADP-ribose) polymerase-1 (PARP-1), and transient receptor potential melastatin 2 (TRPM2) channel levels in heart tissue were measured using an enzyme-linked immunoassay (ELISA) kit. In addition, histopathological examination and immunohistochemical investigation of the TRPM2 channel, fibroblast specific protein-1 (FSP1), transforming growth factor-beta- 1 (TGF-β1) and α-smooth muscle actin (α-SMA) expressions were determined in heart tissue. The CYP group's troponin-I, total cholesterol, triglyceride, CK-MB, ROS, PARP-1 and TRPM2 channel levels were higher than in the other groups in the ELISA measurements (p < 0.05). In contrast, these parameters in the group treated with CURC and BER together with CYP were lower than in the CYP group (p < 0.05). Additionally, CUR and BER reduced CYP-induced pathological damage, TRPM2, FSP1, TGF-β1 and α-SMA expressions. The data showed that CYP administration can cause cardiac damage by increasing the TRPM2 channel, TGF-β1, FSP1 and α-SMA expression levels. Therefore, we concluded that CURC and BER administration following CYP application may be used as therapeutic agents to prevent CYP-induced cardiac damage.

Boswellic Acid and Betulinic Acid Pre-treatments Can Prevent the Nephrotoxicity Caused by Cyclophosphamide Induction.

Cyclophosphamide (CYP) is a chemotherapeutic drug used to treat various cancers. However, its clinical use is limited due to severe organ damage, particularly to the kidneys. While several phytochemicals have been identified as potential therapeutic targets for CYP nephrotoxicity, the nephroprotective effects of boswellic acid (BOSW) and betulinic acid (BET) have not yet been investigated. Our study used 42 rats divided into six equal groups. The study included six groups: control, CYP (200 mg/kg), CYP+BOSW20 (20mg/kg), CYP+BOSW40 (40 mg/kg), CYP+BET20 (20 mg/kg), and CYP+BET40 (40 mg/kg). The pre-treatments with BOSW and BET lasted for 14 days, while the application of cyclophosphamide was performed intraperitoneally only on the 4th day of the study. After the experimental protocol, the animals were sacrificed, and their kidney tissues were isolated. Renal function parameters, histological examination, oxidative stress, and inflammation parameters were assessed both biochemically and at the molecular level in kidney tissue. The results showed that oxidative stress and inflammatory response were increased in the kidney tissue of rats treated with CYP, leading to impaired renal histology and function parameters (p < 0.05). Oral administration of both doses of BET and especially high doses of BOSW improved biochemical, oxidative, and inflammatory parameters significantly (p < 0.05). Histological studies also showed the restoration of normal kidney tissue architecture. BOSW and BET have promising biological activity against CYP-induced nephrotoxicity by attenuating inflammation and oxidative stress and enhancing antioxidant status.

Cyclophosphamide exerts an anti-metastatic effect by reducing the expression of MMP-2 and -9 in Saos2 osteosarcoma cells

Purpose: Although osteosarcoma is the most common primary malignant bone tumor among adolescents and young adults, the molecular mechanisms underlying the development of cancer are not fully elucidated. Cyclophosphamide (CYC) is an alkylating antineoplastic agent widely used in the treatment regimens of various cancers and chronic inflammatory diseases. The aim of this study is to reveal the effect of CYC on Saos-2 cells at the molecular level by determining the expression levels of some matrix metalloprotease proteins (MMP-2 and MMP-9) of cancer progression related genes.&#x0D; Materials and Methods: Cytotoxicity analyzes were performed by applying CYC to the developed human osteosarcoma (Saos-2) cell cultures. The determined dose of CYC was applied to the 2D cell lines for 12, 24 and 48 hours. As a result of agent application, the expression levels of MMPs, which are metastatic markers in these cultures, were determined by immunocytochemical method.&#x0D; Results: While MMP-2 and MMP-9 protein expression levels increased in the control (PBS) groups in correlation with the extend in duration in the cell cultures created; decreased with CYC administration.&#x0D; Conclusion: It was determined that the expression levels of genes that play a role in metastasis such as MMP-2 and MMP-9 were decreased by CYC application to Saos-2 cells. These results shed light on molecular studies for the treatment of osteosarcoma cancer.

Protective effect of myricetin, apigenin, and hesperidin pretreatments on cyclophosphamide-induced immunosuppression

Aim: Major side effects of cyclophosphamide administration are immunosuppression and myelosuppression. The immunomodulatory effects of plant bioactive compounds on chemotherapy drug-induced immunosuppression may have significant effects in cancer treatment. For this reason, we investigated the immunomodulatory effect of myricetin, apigenin, and hesperidin in cyclophosphamide-induced immunosuppression in rats. Methods: In our study, a total of 64 rats were used, and divided into eight equal groups. These groups were: control, cyclophosphamide, cyclophosphamide + myricetin (100 mg/kg), cyclophosphamide + myricetin (200 mg/kg), cyclophosphamide + apigenin (100 mg/kg), cyclophosphamide + apigenin (200 mg/kg), cyclophosphamide + hesperidin (100 mg/kg), and cyclophosphamide + hesperidin (200 mg/kg). Myricetin, apigenin, and hesperidin pretreatments were performed for 14 d, while cyclophosphamide application (200 mg/kg) was performed only on the 4th day of the study. Levels of humoral antibody production, quantitative hemolysis, macrophage phagocytosis, splenic lymphocyte proliferation, and natural killer cell cytotoxicity were determined. In addition, we measured pro-inflammatory cytokines, and followed lipid peroxidation and antioxidant markers and examined the histology of bone marrow, liver and spleen in all groups. Results: During cyclophosphamide treatment, all three phytochemicals increased the levels of humoral antibody production, quantitative hemolysis, macrophage phagocytosis, splenic lymphocyte proliferation, antioxidant markers, and natural killer cell cytotoxicity. Moreover, the agents decreased the levels of pro-inflammatory cytokines and mediators, reduced lipid peroxidation markers, and reduced tissue damage in liver, spleen, and bone marrow. Conclusion: Our study demonstrated that myricetin, apigenin, and hesperidin can reduce the immunosuppressive effect of cyclophosphamide by enhancing both innate and adaptive immune responses, and these compounds may be useful immunomodulatory agents during cancer chemotherapy.

Analysis of Risk Factors of Herpes Zoster in Patients with Multiple Myeloma Treated with Bortezomib

To explore and analyze the risk factors of herpes zoster in patients with multiple myeloma (MM) during the chemotherapy with bortezomib. Clinical data of 85 MM patients treated with bontizomib from January 2015 to January 2019 were selected and divided into case group and control group accroding to the occurred of herpes zoster. The clinical characteristic, treatment outcome and related factor of herpes zoster were retrospective analyzed. Twenty of the 85 patients with MM treated with bortezomib developed herpes zoster occurred (23.5%). Single-factor analysis showed that age≥65 years, lymphocytopenia occurred before treatment, neutropenia occurred before treatment, ECOG score≥2, application of cyclophosphamide, absence of preventive antiviral therapy were associated with the genesis of herpes zoster (P＜0.05). Multivariate logistic regression analysis showed that lymphocytopenia occurred before treatment, the application of cyclophosphamide and the absence of preventive antiviral therapy were the independent risk factors for herpes zoster (P＜0.05). The incidence of herpes zoster is high in the multiple myeloma patients treated with bortezomib. Lymphocytopenia occurred before treatment, the application of cyclophosphamide, and the absence of prophylactic antiviral therapy are the important risk factors for herpes zoster, for which the clinicians should attach great importance.

Systémová zánětlivá reakce s vysokými hodnotami CRP jako dominantní příznak mnohočetného myelomu

A man aged 60 years was examined for intense inflammatory response, night sweats, subfebrile and later febrile temperatures and a weight loss of 18 kg in 7 months. CRP was 270 mg / l, i.e. more than 20 times the upper limit of the physiological range. Reactive leukocytosis (10 × 109/l), thrombocytosis (530 × 109/l), increased fibrinogen (greater than 7 g/l), and anemia with hemoglobin of 80 g/l were present. No infection or systemic autoimmune disease has been proven. The patient had normal renal function and had no osteolytic deposits detectable by FDG-PET/CT. The procalcitonin level was not elevated. The bone marrow examination revealed a 30-40% infiltration of proplasmacyte type with admixture of plasmablasts, expressing light chains λ. Monoclonal immunoglobulin IgA λ was at a low concentration of about 8 g/l and the ratio of free light chains κ/λ was 0.13. The extent of bone marrow infiltration and anemia met the criteria for the diagnosis of symptomatic multiple myeloma. Following initiation of the combination therapy using thalidomide, bortezomib and dexamethasone, the maximum decrease in the concentrations of monoclonal immunoglobulin, free light chains and CRP was observed already after the first 2 cycles of treatment. Later, during the following two 2 cycles, the disease began to progress again. The patient underwent successful stem cell collection after the application of cyclophosphamide 2.5 g/m 2 and leukocyte growth factor (G-CSF), and high-dose chemotherapy (melphalan 200 mg/m 2) with the support of stem cell transplantation. At 2 months following high-dose chemotherapy, CRP levels of the physiological range decreased, the blood count was normalized, and monoclonal immunoglobulin was not detectable. Conclusion: The chronic inflammatory response may be due to plasmocytary bone marrow infiltration even if there are no other symptoms of multiple myel-oma present, except for anemia which, however, also involves the inflammatory reaction. In this case, the systemic inflammatory reaction with high CRP levels signalled aggressive behaviour of the disease. Key words: CRP - multiple myeloma - procalcitonin - systemic inflammatory response.

новая биологическая модель умеренного торможения роста опухоли и метастазов с продолжительной лейкопенией у мышей

A new biological model of moderate inhibition of tumor growth and metastases with prolonged leukopenia on C57Bl/6 mice with the Lewis Lung Carcinoma was designed. The model was created by the injection of cyclophosphamide (dose 83.3mg/kg) on 6th, 12th, 18th days after tumor cells transplantation on animals. Experiment showed that 3-fold cyclo-phosphamide use leads to growth of primary tumor and metastases inhibition. Tumor growth inhibition was 34% on 21st day after cyclophosphamide inject. The number of metastases decreased by 4.7times (p&amp;#60;0,01). Metastatic area reduced. Metastasis frequency made 100%. In addition, the course of cyclophosphamide application caused inhibition of granulocytic and lymphoid hematopoiesis. The reducing the number of segmented neutrophils and lymphocytes was showed on the 3rd day after 1, 2 and 3 injections of cyclophosphamide. The model can be used to study the efficacy of drugs in tumor therapy and in correction of such toxic manifestation of chemotherapy as leukopenia.

Treatment with cyclophosphamide supported by various dendritic cell-based vaccines induces diversification in CD4⁺ T cell response against MC38 colon carcinoma.

The present study shows that an application of cyclophosphamide (CY) supported by dendritic cell (DC)-based vaccines affected differentiation of the activity of CD4+ T cell subpopulations accompanied by an alteration in CD8+ cell number. Vaccines were composed of bone marrow-derived DCs activated with tumor cell lysate (BM-DC/TAgTNF-α) and/or genetically modified DCs of JAWS II line (JAWS II/ Neo or JAWS II/IL-2 cells). Compared to untreated or CY-treated mice, the combined treatment of MC38 colon carcinoma-bearing mice resulted in significant tumor growth inhibition associated with an increase in influx of CD4+ and CD8+ T cells into tumor tissue. Whereas, the division of these cell population in spleen was not observed. Depending on the nature of DC-based vaccines and number of their applications, both tumor infiltrating cells and spleen cells were able to produce various amount of IFN-γ, IL-4 and IL-10 after mitogenic ex vivo stimulation. The administration of CY followed by BM-DC/TAgTNF-α and genetically modified JAWS II cells, increased the percentage of CD4+T-bet+ and CD4+GATA3+ cells and decreased the percentage of CD4+RORγt+ and CD4+FoxP3+ lymphocytes. However, the most intensive response against tumor was noted after the ternary treatment with CY + BM-DC/TAgTNF-α + JAWS II/IL-2 cells. Thus, the administration of various DC-based vaccines was responsible for generation of the diversified antitumor response. These findings demonstrate that the determination of the size of particular CD4+ T cell subpopulations may become a prognostic factor and be the basis for future development of anticancer therapy.

Effect of IEPA, a Novel Orally Bioavailable Small Molecule, on Chemotherapy-Induced Myelosuppression

Introduction: Leucopenia following myelosuppressive chemotherapy is associated with substantial mortality and costs. Whereas there is a lack of therapeutic options for lymphopenia, in chemotherapy-induced neutropenia, Granulocyte Colony-Stimulating Factor (G-CSF) has proven to be effective in secondary prophylaxis. Despite currently available therapies, mortality due to chemotherapy-induced neutropenia remains high, and safety, tolerability and costs limit the use of available treatment options.Imidazolyl Ethanamide Pentandioic Acid (IEPA, Myelo001) is a novel small molecule for the treatment of chemotherapy-induced leucopenia. Preclinical and clinical studies have shown that in its oral form IEPA is effective in reducing neutropenia and has antiviral properties without noticeable toxicity.Objective: To evaluate the effect of IEPA after multiple oral administration in different posologies and at a low (1.0mg/kg) and high dose (100mg/kg) on peripheral components of hematopoiesis in a mouse model of acute cytostatic myelosuppression in comparison to vehicle and Filgrastim (Neupogen¨).Methods: Twenty-five female CD-1 mice, 6 weeks of age, were randomly assigned to 5 treatment groups: Cyclophosphamide (CPH) (200mg/kg), CPH+Filgrastim (subcutaneous) (0.3mg/kg) (quaque die (q.d.) day (d) 1 to 5), CPH+IEPA (100mg/kg) (q.d., d -1 to 5), CPH+IEPA (100mg/kg) (q.d., d -5 to 5), CPH+IEPA (1.0mg/kg) (q.d., d -5 to 5). The experimental phase started with the determination of baseline white blood cell, red blood cell and platelet counts one day before (pre-) treatment start with IEPA (d -6) and was continued on days 1, 3, 5, 8, 23, 32. A single intraperitoneal (i.p.) injection of Cyclophosphamide (200mg/kg) was administered in all 5 groups on day 0 to induce leuco- and thrombocytopenia. The longitudinal data were analyzed by response profile analysis.1Results: The three groups of IEPA had lower baseline leucocyte counts compared to the CPH+Filgrastim and CPH groups. CPH application caused leucopenia across all 5 groups with the nadir on day 3 to 5. Comparing change from baseline, IEPA significantly reduced leucopenia on days 3, 5, 8 relative to the control group (CPH). No clear dose- or posology-effect relationship was discernible between doses of 1.0 (for 11d), 100 (7d) or 100mg/kg (11d). Compared to Filgrastim, IEPA led to an earlier recovery of leucocytes, followed by a delayed and less pronounced increase in leucocytes above baseline, with the peak of Filgrastim (change of baseline relative to CPH) on day 5 (10.2∙109/L, 95%CI: 7.1 to 13.2∙109/L, p < 0.001) versus day 8 for IEPA (change of baseline relative to CPH) (4.7∙109/L, 95%CI: 1.5 to 8.0∙109/L, p = 0.004 at a dose of 100mg/kg (11d)) [Display omitted] [Display omitted] Lymphocyte counts change from baseline in all three IEPA groups was numerically higher than in the Filgrastim and CPH arm on day 3, 5 and 8, and day 3 and 8, respectively (not statistically significant). Filgrastim led to a substantial granulocytosis on day 5 (change of baseline relative to CPH) (7.2∙109/L, 95%CI: 5.6 to 8.9∙109/L p < 0.001), whereas a minor increase was observed in the three IEPA groups on day 8. Thrombocytes numbers were slightly increased throughout days 3 to 23 in the IEPA groups, contrasting with a pronounced thrombocytopenia under Filgrastim on day 5. Red blood cell counts decreased after CPH injection (nadir on day 3 to 8), but did not differ among groups.Conclusion: IEPA demonstrated in 2 doses and posologies a consistent reduction of chemotherapy-induced leucopenia and thrombocytopenia in CD1 mice. Its effect on leucopoiesis and thrombocytopoiesis differs from Filgrastim in a less pronounced early nadir and subsequent lower amplitude of leuco- and granulocytosis. IEPA may offer a new therapeutic option for myelosuppression due to chemotherapy, but requires further preclinical and clinical investigation.1Fitzmaurice GM, Laird NM, Ware JH. Applied longitudinal analysis, Wiley. Hoboken, NJ: Interscience; 2004 p103-140. DisclosuresPleimes:Myelo Therapeutics: Employment, Equity Ownership, Managing Director Other; Bayer Pharma AG/ Bayer Healthcare Pharmaceuticals: Consultancy. Flechsig:EPO Berlin-Buch GmbH: Contract Research on behalf and in account of Myelo Therapeutics GmbH Other, Employment. Meyer:Myelo Therapeutics GmbH: Consultancy.

Cyclophosphamide and IL-12-transduced DCs enhance the antitumor activity of tumor antigen-stimulated DCs and reduce Tregs and MDSCs number.

A hostile tumor microenvironment, characterized by an abundance of T regulatory cells and myeloid-derived suppressor cells (MDSCs), considerably limits the efficacy of dendritic cell (DC)-based vaccines. The intention of this study was to enhance the antitumor activity of vaccines consisting of bone marrow-derived DCs stimulated with TAg (BMDC/TAg) via single administration of cyclophosphamide and multiple injections of interleukin (IL)-12-transduced DCs (BMDC/IL-12). The combined chemoimmunotherapy was applied in the treatment of mice with subcutaneously (SC) growing, advanced MC38 colon carcinoma. The highest level of tumor growth inhibition, accompanied by high cytotoxic activity of effector cells, and their increased influx into tumor tissue, was observed after application of cyclophosphamide in combination with BMDC/TAg and BMDC/IL-12. The effect was probably associated with the elimination of T regulatory cells from spleens and tumors, but most of all with changes in the number and differentiation stage of MDSCs. After the therapy, the percentage of granulocytic and monocytic MDSCs in spleens was significantly lower than in the control group. Moreover, MDSCs derived from spleens and tumors showed increased expression of MHC class II, which may indicate the higher maturation stage of the myeloid cells as well as their enhanced capacity toward antigen presentation. The obtained data indicate that the optimal composition of antitumor vaccines able to limit the suppressor activity of MDSCs is essential to enhance the elimination of tumor cells and to achieve an optimal therapeutic effect.

Prolonged and Devastating Clinical Course in Atypical Neuromyelitis Optica

Neuromyelitis optica (NMO) is an inflammatory demyelinating disease affecting the optic nerve and the spinal cord. Antibodies (Ab) to Aquaporin-4 (AQP-4) and sometimes to myelin oligodendrocyte glycoprotein (MOG) are characteristic serological findings. We present the case of a 9-year-old boy with seronegative atypical NMO showing recurrent episodes of isolated longitudinal extensive transverse myelitis (LETM). He presented in December 2010 with paresis of his left leg following flu vaccination. LETM with areas of beginning necrosis was diagnosed. Blood-brain barrier disruption, oligoclonal band and highly elevated interleukin 6 were found in cerebrospinal fluid. Clinically, the entire spinal cord was affected with tetraparesis and inverted breathing. He was treated with steroids, plasmapheresis, cyclophosphamide (CPM), and immunoglobulins. Paraplegia starting from Th9 and a neurogenic bladder disorder persisted. AQP4- and MOG-Ab were repeatedly tested negative. There was no evidence of optic neuritis during the follow-up period. In July 2013, he experienced a first relapse with weakness of the upper extremities following oral herpes infection. CSF, blood, and magnetic resonance images showed identical findings to the first episode. After steroids, plasmapheresis and CPM he showed no clinical improvement and lost his hand function. After 27 days, application of CPM again following oral herpes infection he experienced a second relapse with tetraparesis, sensibility loss starting at Th2, dry mouth, and neck pain. Lumbar puncture was no longer possible. He showed no improvement to steroids and plasmapheresis, but the inflammation progressed to the midbrain. Finally, respiratory failure, seizures, and cardiac arrest September 2013. In conclusion, we suspect NMO-spectrum disorder (high-risk NMO), possibly triggered by infection or vaccination without NMO- and MOG-Ab nor optic nerve involvement. The clinical picture is characterized by a polyphasic course with late relapses and poor prognosis. Our case demonstrates an unusual clinical course of LETM and should open discussion about the variable clinical picture of NMO in childhood.

613. Challenging an Old Drug in a New Context: Low-Dose Cyclophosphamide in Immunogenetherapy of Cancer

Top of pageAbstract Breaking down immunological tolerance towards cancer cells is a major hurdle towards the aim of effective immunotherapy. Naturally occurring CD4+CD25+ T regulatory cells (Tregs) have an important role in peripheral self-tolerance and recent reports indicate that Tregs also mediate immunotolerance towards tumors. Cyclophoshamide (Cy) has been approved as a therapeutic drug since the 1970s. High- dose cyclophosphamide application is used for immunosuppression and cancer therapy due to its cytostatic and anti-angiogenetic effects. In contrast low-dose Cy application has been shown to trigger immune responses against cutaneous contact allergens and tumor- antigens. Here, we investigate the effect of low-dose Cy treatment on immune and cancer cells in a mouse model. MATERIAL AND METHODS: Immunocompetent and immunodeficient (SCID) C57Bl/6 mice received a single i.p. injection of Cy ranging from high-dose (200mg/kg) to low-dose (50mg/kg) and were sacrificed at different time-points for analysis of normal, malignant and immunecells. Syngenic TC-1 and EL-4 cells served as tumor models. To assess the effect of Cy on immunogenetherapy, adenoviral vectors expressing anti-CD3 and fetal liver tyrosine kinase 3 (Flt3 ligand) were constructed and applied intratumorally. RESULTS: High-dose Cy application led to a general immunosuppression in immunocompetent mice and delayed tumor growth in both immunocompetent and immunodeficient mice. In contrast, low-dose Cy strongly delayed tumor growth in immunocompetent, but not in immunodeficient mice. When Cy treatment was combined with immunotherapy, using the vectors described above, tumors were eliminated, whereas application of the vectors alone had no curative effect. Re-challenging with tumor cells led to delayed tumor growth when compared to naive mice. Immunohistochemistry analysis showed infiltration of immune-effector cells into the tumors upon low-dose Cy injection. Intracellular cytokine staining revealed activation of tumor-specific cytotoxic CD8+ T-cells. Our data suggest that low-dose Cy has a preferential effect against Tregs, whereas immune effector cells (e.g. CD4+ and CD8+ T-cells) are less Cy sensitive. Low-dose Cy depletes Tregs at the tumor site and in secondary lymphoid organs. As a serious side effect low-dose Cy downregulates CD19+ B-cells. Both, the number of splenic Tregs and CD19+ cells declined by day 4 after low-dose Cy injection and returned to normal levels by day 7. No signs of autoimmune diseases in low-dose Cy treated mice were observed. CONCLUSIONS: We show, that a single application of low-dose Cy is capable of inducing a strong immune response against autologous tumor cells. Moreover the susceptibility of tumors towards adenoviral immunogene- transfer was significantly increased leading to immune-mediated eradication of large tumors. Our data provide a rationale for using low-dose Cy in immunotherapy of cancer, either as a single agent or in combination with immunogenetherapy approaches.

Effects of a modified CMF treatment (cyclophosphamide, methotrexate and 5-fluorouracil) on hematopoietic tissues and Yoshida sarcoma in rats.

Effects of a modified CMF treatment on hematopoietic tissue and an implanted tumor were studied in rats. The modification of the treatment refers to the application of cyclophosphamide 24 h after methotrexate and 5-fluorouracil. The study was done on Wistar rats bearing Yoshida sarcoma in the ascites form. The controls were a) untreated animals bearing the tumor or b) treated conventionally with the 3 cytostatics and c) tumor-free animals under either conventional treatment or d) modified treatment. We examined survival, the appearance of metastases, and the regeneration of hematopoietic tissues. Improved survival, the absence of metastases, and improved regeneration of hematopoietic tissues was observed when modified CMF treatment was applied. These results support the importance of sequencing cytostatic protocols for basic hematological determinants and anti-tumor activity.

Effect of different local and systemic therapy upon urinary bladder cytology.

1,900 cytological analyses of urine and bladder washings were made in 127 patients with urothelial bladder carcinomas before, during and after therapy. Following transurethral resection, all patients were treated by intravesical instillation of mitomycin C or thiotepa. Because of a locally advanced bladder carcinoma, 26 patients who were not candidates for radical cystectomy were given an integrated treatment of radiotherapy and chemotherapy. Intravesical administration of mitomycin C and thiotepa as well as integrated radiotherapy and chemotherapy induce a variety of cytological effects (toxic and/or metabolical) which may lead to cytological misinterpretations in the follow-up. DNA measurements by means of single-cell spectrocytophotometry show that the cytological effects induced by the above-mentioned therapies are not accompanied by an increase in the nuclear DNA content. It is concluded that the knowledge of these induced effects is mandatory for a correct interpretation of urinary cytology in the follow-up. Considering these effects and the clinical history, bladder carcinoma recurrences during and after intravesical chemotherapy or integrated radiotherapy and chemotherapy may be detected early by urinary cytology in the hands of an experienced cytopathologist or urologist. Furthermore, alterations of the urinary cytology occur after systemic application of cyclophosphamide and under immunosuppressive therapy.

Zur Normogenese der Hornhaut und ihrer Beeinflussung durch Cyclophosphamid im Tierexperiment

Animal experiments serving as models have hitherto been indispensable in order to explain pathogenetic processes in the development of malformations. The results presented here are contribution to studies on the normogenesis of the cornea in defined animal material (albino mice of Agnes Bluhm Jena-Halle stock). A single application of cyclophosphamide during embryogenesis caused inhibition malformations. Of these, different degrees of corneal hypoplasia and Peters' malformation are discussed in detail.

In vitro/in vivo effects of Mesna on the genotoxicity and toxicity of cyclophosphamide — a study aimed at clarifying the mechanism of Mesna's anticarcinogenic activity

Cyclophosphamide is an effective antitumor agent with considerable side effects such as urotoxicity and carcinogenicity. These negative attributes may be caused by toxic and genotoxic metabolites, respectively. Mesna (sodium 2-mercaptoethane sulfonate) decreases the urotoxieity by scavenging the toxic metabolite acrolein. The present study was aimed at elucidating whether a similar scavenging of genotoxic alkylating intermediates could be found, which might cause the reduction in carcinogenicity. In vitro studies on the genotoxic and toxic properties of cyclophosphamide and its major metabolites to bacteria were therefore performed in the presence of Mesna. Mesna did not reduce the mutagenicity of any of the tested metabolites. Mesna clearly inhibited the toxic properties of acrolein. After in vivo application of Mesna and cyclophosphamide to rats, however, a lower yield of mutagens in the excreted urine was observed than after application of cyclophosphamide only.

Metallophilic and PAS‐positive Cells of the Rat Thymus, Topographical Distribution and Histological Properties after the Application of Cyclophosphamide

AbstractThe topographical distribution, number and histological properties of metallophilic and PAS‐positive cells were investigated during the process of spontaneous regeneration of the thymic tissue after the application of consecutive doses of Cyclophosphamide (5 ± 60 mg/kg). The aim of the study was to elucidate the relationship between these cell types which are both localized in the cortico‐medullary zone of the normal rat thymus.

Restriction of regenerative capacity of the rat thymus after the application of cyclophosphamide

Restriction of thymic regenerative capacity, in contrast to the data in the literature, was found 30 days after a single (300 mg per kg) or fractionated doses (5 × 60 mg per kg) of Cyclophosphamide. Detailed cellular analysis revealed myelometaplastic changes in the thymic parenchyma, especially in the outer cortex and cortico-medullary zone. Plasmoid elements also appeared in the outer cortex. The capsular and septal connective tissue was markedly thickened. The thymic mass was decreased. The changes were more marked after the fractionated than after the single dose of Cyclophosphamide.

Abnormal base pairing under the influence of nitrogen mustard

Cyclophosphamide, frequently used in tumor therapy, has proved to be mutagenic [l-4], but so far no clear evidence has been obtained in the host-mediated assay as to whether the substance causes point mutations [5]. Therefore we decided to approach the problem biochemically. Ludlum [6,7] has recently shown that the methylation of cytidine-diphosphate, or poly(C), and to a lesser degree that of the corresponding deoxyderivatives, alters the pairing characteristics of this base in such a way that in the transcription and/or polymerization process not only guanine, but also though to a smaller extent uracil and adenine are incorporated into the resulting polymer. Similar observations regarding the misreading of chemically altered poly(C) have been made by Singer and FraenkelConrat [8,9] and also by Means and Fraenkel-Conrat [IO]. On the basis of the results they obtained with several mutagens, these authors suggest that the substitution of position 3 of cytosine by an alkyl group might well represent a mutagenic event, inasmuch as methylated poly(C) shows appreciable ambiguity, thus leading to abnormal base pairing (with simultaneously diminished capacity for normal pairing with guanine). Since cyclophosphamide as such is not an alkylating agent, we utilized 0, /I’-dichlordiethylamine (nornitrogen mustard) (I) in our experiments. This compound has been postulated to be one of the active substances formed in the body after application of cyclophosphamide [ 111. Therefore, because of the alkylating and cross-linking properties of I [ 121, a strong diminution in the template function of poly(C) after incubation with I is by no means surprising. What could not be foreseen, however, was whether the treatment of poly(C) with such a relatively large molecule (large in comparison with methyl groups) would lead to abnormal base pairing. In fact, our findings have shown a very slight tendency towards the incorporation of LIMP, significantly higher than the “spontaneous” rate of abnormal base pairing with UMP, and increasing linearly with the time of incubation.

KOYUNLARIN İLAÇLI KIRKIMLARI VE BU AMAÇLA MERİNOSLAR ÜZERİNDE YAPILMIŞ OLAN BİR CYCLOPHOSPHAMIDE UYGULAMASI

OZET Bir yasindaki erkek Merinoslar uzerinde, ilacli kirkimdaki etkisini kontrol etmek amaci ile Cyclophosphamide uygulamasi yapilmistir. ilacin etki zamani ile yapagi verimi ve yapaginin yolunup alinabilmesi icin gerekli olan zaman, kontrol gurubu ile karsilastirmali olarak, incelenmis ve bulunmus olan sonuclarin bu konudaki literatur ile uyarlilik' halinde olduklari gorulmustur. CHEMICAL SHEARING OF SHEEP AND A CYCLOPHOSPHAMIDE APPLICATION ON MERlNO RAMS 1". In this investigation the Cyc1ophosphamide was used to sheare the .Merino Rams under Erzurum conditions. 2. 56 Merino Rams, one year old, were used in this study. They were randomly divided into two groups. One of the groups was used for the drug administration, and the other one was kept as a control group and sheared mechanka1ly at the same time with the former. 3. Both of the groups were kept 'under similar conditions ~md they were never shorn before. 4. A 4 % solution of CyCiophosphamide was given to animals orally, on the basis of 25 mg./kg. boCiy weight. The experiInetal animals were no treated spesifically before the administration of the drug. 5. Up to sixth day, after the administration of th~ drug, animals shO'\>J'ed no clinical signs. 6· At the contrals done by pulling of fleece 10cally, at the sixth day of the dosing of animals, the skin was found completely bare underneath. For this reason to remove the fleece it was waited ti11 to 2] st. days, as the literature stated A:fter that time all the fleece were Temoved from the animals. Animals body came out with short, unuorm newgrowth hair. 7. The fleece could not be removed from the six of the 28 animals. In the animals the time spent for removing was 'different amQng animals. Inspite of this fact the averag~ time spent for chemical shearing was shorter 'than the mechanical shearing. It was conc1uded that 30 mg. drug for per kilogram body is necessary t9 eliininate this variance which might come from the indidividual. 8, At the comparison of the fleece obtained fro~ both groups, the average weight of fleece was greater in the drug administred _group than the mechanically shom group. Because therc was not any shearing loss. in the former. 9. General1y it can be stated that the method of chemical shearing could ve replaced widely the mechanical sheaong in the very future. Bu it would be necessary that: a. The drug must be prepared so in a form that the animal raiser can dose it easely, b. The cost of the drug apHed per animaI must be cheaper than the cost of mechanical shearing, c. And much research must be done on the side ~ffects of the drug to be sure that it is not harmfull to anima1s and also to human health at alL.