Chemotherapy for mobilisation of Ph-negative progenitor cells from patients with CML: impact of different mobilisation regimens.

Mobilised peripheral blood stem cells are widely used for autografting in patients with chronic myeloid leukaemia (CML) and it is generally thought that a high proportion of Ph-negative progenitor cells in the graft is desirable. We report here the results of 91 stem cell mobilisations performed with various chemotherapy regimens followed by G-CSF. We show that mobilisation of Ph-negative cells is possible after diagnosis as well as in advanced stages of the disease. The yield of Ph-negative cells is highly dependent on the chemotherapy regimen: while the combination of idarubicin and cytarabin for 3-5 days (IC3-5) mobilised Ph-negative cells in most patients, high-dose cyclophosphamide was ineffective. Mobilisation of Ph-negative progenitor cells after IC3 was at least as effective as after IC5; however, less apheresis sessions were required, and toxicity was much reduced after IC3. Compared to historical controls, IC was equally effective as the widely used ICE/miniICE (idarubicin, cytarabin, etoposide) protocol. No correlation was found between graft quality and the cytogenetic response to subsequent treatment with interferon-alpha. We conclude that IC3 is an effective and well-tolerated regimen for mobilising Ph-negative cells that compares well with more aggressive approaches such as IC5 and ICE/miniICE.

Tolerance to the antinociceptive effect of Crotalus durissus terrificus snake venom in mice is mediated by pharmacodynamic mechanisms.

Crotalus durissus terrificus venom exerts central and peripheral antinociceptive effect mediated by opioid receptors. The present work investigated the tolerance to the antinociceptive effect of the venom and characterised the mechanisms involved in this phenomenon. The hot plate test, applied in mice, was used for pain threshold determination. The venom (200 microg/kg) was administered by oral route, daily, for 14 days, and the nociceptive test was applied before and on days 1, 7 and 14 of the treatment. Prolonged treatment with venom lead to the development of tolerance to the antinociceptive effect. Tolerant animals exhibited increased sodium pentobarbital-induced sleeping time, although total hepatic microsomal cytochrome P450 was not altered. The antinociceptive effect of a single dose of venom (200 microg/kg) is mediated by kappa opioid receptors. Mice long-term-treated with venom showed cross-tolerance to U-TRANS, an agonist of kappa-opioid receptor, but not to morphine or DAMGO, two mu-opioid receptor agonists. Prolonged administration of venom did not cause symptoms of abstinence syndrome. These data indicate that prolonged treatment with C. durissus terrificus venom induces tolerance to the antinociceptive effect and that pharmacodynamic mechanisms are involved in the genesis of this phenomenon.

[Serological diagnosis of cytomegalovirus-induced interstitial pneumonia in patients following bone marrow transplantation]. / Serologische Diagnostik zytomegalievirus-bedingter interstitieller Pneumonien bei Patienten nach Knochenmarktransplantation.

For diagnosis of the interstitial pneumonia conditioned by cytomegalovirus (CMV) an indirect immunofluorescence test was used to measure IgM and IgG antibodies to CMV in 369 serum specimens from 41 patients after bone marrow transplantation (BMT). An interstitial pneumonia conditioned by CMV was diagnosed in 5 patients either serological (4 patients) or by detection of cytomegaloviral infection typical inclusion bodies in lung cells (1 patient). The detection of antibodies to CMV for diagnosis of interstitial pneumonia conditioned by CMV is problematical in the phase immediately after BMT because bone marrow recipients are severely immunosuppressed by radiation and cytotoxic drugs received before transplantation. New method for detection of CMV-specific antigens are necessary.

Allogeneic bone marrow transplantation for leukaemia and aplastic anaemia. Results of the Leipzig BMT Group.

In acute leukaemias there was a stable plateau in the survival curve at 45% after two years if grafted in first complete remission (n = 20) but only 13% of the patients are disease-free alive if grafted in a more advanced stage of the disease (n = 8). In 16 patients transplanted for chronic myeloid leukaemia the overall survival is 40%, in cases with graft-versus-host disease (GVHD) prevention by cyclosporine survival rate could be improved. Only 8 patients with severe aplastic anaemia, partially in low performance status were able to be transplanted; three died of infections, another by acute GVHD. The fatal complications in our study characterize the international well-known major problems in BMT: GVHD, interstitial pneumonitis, infections, graft failure in aplastic anaemia and recurrence of leukaemia, especially in more advanced leukaemia stage.

Acute and late effects in the course of and after total body irradiation with following bone marrow transplantation in patients with different forms of leukaemia.

The acute reaction in the course of a total body irradiation (TBI) appears in an organ-specific damage of the stem cells. Moreover, there are unspecified central-nervous stress reactions. Clinical reactions are obtained by the study and symptomatic therapy is proposed. 90 patients with different forms of leukaemia were observed. We documented the course in a specific "protocol system". Reactions like an increase of body temperature, changes of pulse and blood pressure were registered. The occurrence of gastro-intestinal reactions is a typical symptom of the acute radiation syndrome e.g. vomiting and diarrhoea are demonstrated in dependence of the applicated dose of irradiation. Further symptoms of TBI appeared in the later period. Mucositis, parotitis, a decreased function of the salivary glands and diarrhoea as well as vomiting are characterized by different intensity and temporary termination. A difference between allogeneic and autologous transplantation is caused by a medicamental additional treatment. During the late period these symptoms will disappear completely. Moreover, after TBI and BMT late effects are a cataract and some changes in the hormonal system demanding a specific correction or substitution respectively.

Autologous bone marrow transplantation (ABMT) using unpurged marrow as intensification for first complete remission in acute leukaemia (AL).

In order to assess the clinical advantage of autologous bone marrow transplantation (ABMT) without ex vivo purging, the results in 26 patients (10 AML, 16 ALL) in 1. CR were analyzed retrospectively. All patients received 3 consolidation cycles "in-vivo purging" before marrow harvesting. Beside relapses infections and cardiac failure were the most frequent complications. After 1 to 12.5 months 11 cases relapsed with a higher probability in patients who had a longer period of induction and between CR and ABMT. 12 patients became relapse-free survivors 6 to 53 months after ABMT with a stable plateau after 12.5 months for 8 patients. In conclusion, ABMT following "in-vivo purging" as the strongest one-step postremission therapy in patients with acute leukaemias may be a way for better long-term results in these patients.

Endocrine dysfunction after total body irradiation and bone marrow transplantation.

The present report describes our data regarding changes of endocrine parameters after total body irradiation (TBI) and bone marrow transplantation (BMT). Endocrine glands are usually resistant to irradiation under morphological aspects. But new methods of determination and sensitive tests were developed in the last few years. Now it is possible to detect already small functional changes. Endocrine studies in the course of the disease were followed serially at 16 patients with TBI and BMT. Pretransplant conditioning consisted of single-dose irradiation combined with a high-dose, short-term chemotherapy. Reactions of the endocrine system showed a defined temporary order. Changes of ACTH and cortisol were in the beginning. The pituitary-adrenal cortex system responds in a different way. The pituitary-thyroid system develop a short-term "low-T3-syndrome" reflecting the extreme stress of the organism. At the same time we obtained an increase of thyroxine. Testosterone and luteotropic hormone, the sexual steroids showed levels representing a primary gonadal insufficiency. The studies in the posttransplant period yielded a return to the normal range at most of the hormonal levels with the exception of the sexual steroids. Sterility is one of the late effects of TBI. A tendency towards hypothyroidism could be noticed in some cases being only subclinical forms. Reasons and possible therapy are discussed.

Bacteriological findings in patients with bone marrow transplantation (Karl Marx University Leipzig, 1985-1987).

The results of the bacteriological surveillance cultures for 26 patients with bone marrow transplantation (Karl Marx University Leipzig, G.D.R., 1985-1987) are presented. 5.9% of all surveillance cultures contained facultatively pathogenic germs (with Pseudomonas aeruginosa as the most frequent representative, which was the reason of a sepsis in two patients). Coagulasenegative Staphylococci and other germs with an obscure pathogenicity were isolated upon a large scale, especially from the mucous membrane regions. There are hints, that above all special strains of coagulasenegative Staphylococci "colonize" the patient's body (also for longer periods) and turn into the blood too. During the total decontamination intestinal anaerobic flora is absent. After closing of total decontamination Clostridium perfringens is the first detectable anaerobic species. During the selective decontamination systemic applications of antibiotics are able to obliterate anaerobic findings for certain periods. Recommendations for an effective arrangement of the surveillance cultures of bone marrow transplantation patients are given.

Germ monitoring of patients, staff and environment in reverse isolation.

To estimate the efficiency of germ-poor for patients who underwent BMT or ABMT in reverse isolation we investigated the staff, patients and the environment of the BMT unit bacteriologically and mycologically between 1/85 to 3/87. On patient's body surface gram-negative germs were not estimated but coagulase negative staphylococci persisted. Only in few cases aerobic spores were detectable, seldom enterococci, corynebacterium and fungi. Staff members had a distinct higher grade of contamination inside the BMT-unit but outside of isolation rooms. We found sporadically staphylococcus aureus and germs of the mouth cavity on hands and fore head. Although gram negative rods were seldom detected in environmental studies, humid regions of the ward remain to be problematic zones.

Lethal complications in 30 patients after allogeneic BMT (BMT) and autografted (ABMT) patients (1980-1986) examined by autopsy.

Lethal and accompanying complications from 29 allogeneic, 8 autologous, and 1 syngeneic transplanted pts., died between 1980 and 1986 at the BMT-Center Leipzig were revisited. An important problem was that of systemic and local infections. We observed a predominance of bacterial and fungal infections within the first 3 weeks p.t., and later on a period of predominantly virus-associated infections. The most common early death (up to 3 months p.t.) was related to ICP, as a rule caused by CMV. GvHD was a common finding at autopsy, and GvHD of the bowel seems to facilitate infectious inflammations of the bowel. There was a positive correlation between severe GvHD and the occurrence of ICP. Neither of the died BMT pts. showed signs of relapse. After ABMT ICP did not occur. The lethal complications were septicaemia, local infections, relapse and haemorrhagic diathesis. A toxic effect of DMSO in 3 pts. must be taken into account. In conclusion, a rather small number of lethal factors indicates the steps in improving results of transplantation in the future.

[Results of therapy of 100 patients with acute leukemia--a retrospective unicenter 5-year analysis]. / Therapieergebnisse bei 100 Patienten mit akuter Leukämie--eine retrospektive unizentrische 5-Jahres-Analyse.

The results of therapy in 100 patients who newly fell ill (68 AML, 32 ALL) with acute leukaemia were evaluated (1981 to 1985). The 5-year-survival chance of all patients is 15% for AML, 18% for ALL, first of all it is depending on the degree of remission obtained. The CR rate is nearly 43% (AML) and 66% (ALL), respectively, shows a dependence upon age and is impaired above all by a high early death rate (supportive therapy). With increasing aggressiveness the results of the remission induction therapy improve, as it becomes clear in a comparison with an evaluation of patients 1965-1980 (CR: 15-32%). Also in the postremission therapy the results of intensive forms of therapy are more favourable: 4 years recurrence-free survival after CR in autologous bone marrow transplantation 50%, in allogenic bone marrow transplantation 40%, in cyclic chemotherapy 17%, in oral permanent therapy 0%. Starting from these findings the present conception of the therapy of acute leukaemias is discussed in connection with the literature.

[Changes in selected parameters of carbohydrate metabolism during short term hyperglycemia therapy in Jensen's sarcoma]. / Das Verhalten ausgewählter Parameter des Kohlenhydratstoff-wechsels unter Kurzzeithyperglykämie beim Jensen-Sarkom.

Biochemical investigations of short term hyperglycemia together with tumor-lysing clostridia were undertaken in order to optimize the use of this combined anti-tumor therapy model. Repeated intraperitoneal glucose injections resulted in an increased concentration of glucose in tumor tissue which lasted several hours and paralleled the blood sugar level in tumor bearing animals. At the same time a tumor dependent deterioration in glucose tolerance appeared likely. Characteristic changes in the intratumor lactate concentration were demonstrable. Tumor acidosis lasted 4 to 7 hours. Tumor pyruvate concentrations varied little with hyperglycemia. Simultaneous blood lactate and pyruvate levels did not differ from those of controls. LDH activity as measured in the tumor and in blood serum gave no clear indication of tumor cell damage.

[Morphological studies on Jensen sarcoma in the rat under artificial short-term hyperglycemia]. / Morphologische Untersuchungen am Jensen-Sarkom der Ratte unter artifizieller Kurzzeithyperglykämie.

Possible actions of artificial short-time hyperglycaemia, which is tested as an adjunctive measure in various therapeutical concepts of tumours (for instance in combination with oncolytic apathogenic Clostridia), were checked by histomorphological investigations in Jensen sarcoma of rats. The glucose-induced hyperglycaemia caused a considerable increase of the extend of necrosis in the whole tumour. These morphological findings gave evidence of the high tumour selectivity of the provable effects of artificial short-time hyperglycaemia.

Acute iron overload in mice: pathogenesis of Salmonella typhimurium infection.

The bacterial growth in the tissues of C3D2F1 male mice was measured during an experimental infection with two Salmonella typhimurium strains (high virulence, strain 2386/74; low virulence, strain L15403). This experimental model was used for evaluation of the pathogenesis in normal and iron-overloaded animals. Acute iron overload was accomplished by intramuscular injections of chelated iron (with 2,3-dihydroxybenzoic acid and citrate) with a single dose of 100 micrograms of iron per mouse. Bacteria were given intraperitoneally 1 h after the iron injection. Serum iron levels, transferrin levels, and the bacteria counts in blood and liver were measured simultaneously in all animals. There was a significant increase of bacterial growth in all tissues in the iron-treated animals. Iron abolished the normal clearance of the bacteria with low virulence from the blood. This study demonstrates that a general iron overload, as determined by an increased serum iron level, resulting from preinjection of iron, enhances bacterial growth.

A milker for mice.

A new apparatus for milking mice was used to obtain milk from four anesthetized mice simultaneously over a period of 15 minutes. By using silicon rubber molded teat cups, eight teats could be milked automatically from each mouse. The average yield was 1.5 ml per mouse. This method was especially applicable to production of large amounts of mouse milk. Milking of rats was achieved with the apparatus without any changes.

[Prognostic significance of histobiopsy findings in bone marrow of malignant lymphomas in adults]. / Zur prognostischen Bedeutung histobioptischer Knochenmarkbefunde bei malignen Lymphomen Erwachsener.

Issuing from the fact that the histobiopsy of the bone marrow is the most sufficient method for the proof of infiltrates of malignant lymphomas in the marrow, in 41 patients with Hodgkin's disease and 89 patients with malignant non-Hodgkin-lymphomas was tested whether already may be rendered prognostic conclusions from the histologic findings of the marrow. In these cases could be shown by the different course in patients with Hodgkin's disease in stage IV with and without affection of the marrow that a specific infiltration of the marrow is accompanied by a significant deterioration of the prognosis, which should be taken into consideration in the therapy planning. In the malignant non-Hodgkin-lymphomas those with a high degree of malignity showed a significantly worse prognosis than those with a low degree of malignity. Thus in the individual case the coordination of a lymphoma to one of these groups of malignity which is possible at the histobiopsy of the iliac crest may already be of prognostic significance also without consideration of the type of lymphoma. The prognostic significance of the type of lymphoma itself could be confirmed by the different course of the survival curves in the histologic subtypes of the immunocytoma. Since finally patients with a lymphoplasmocytic immunocytoma in diffuse affection of the marrow show a significantly worse prognosis than in the focal affection may be concluded that apart from the degree of malignity and the type of lymphoma also the type of affection of the marrow which is to be established histologically seems to have a decisive prognostic significance from which the necessity of a different therapeutic approach may be derived. Further examinations on larger numbers of patients including multivariant analyses will yield still more evident results.