Immunotherapy of advanced renal cell cancer using subcutaneous recombinant interleukin-2 and interferon-α

Summary The combined administration of subcutaneous recombinant human interleukin-2 (rIL-2) and interferon- (rIFN-) was studied in a phase II trial on patients with advanced progressive renal cell cancer. Safety, tolerance and clinical response rate of this outpatient treatment protocol were assessed in 29 evaluable patients who received a total of 47 cycles, each consisting of s.c. rIL-2 at 14.4–18 million IU m^–2day^–1 on days 1 and 2, followed by 6 weeks of combined administration of s.c. rIL-2 at 3.6–4.8 million IU m^–2 day^–1 on 5 days a week, and s.c. rIFN- at 3–6 million units m^–2 three times weekly over a period of 6 consecutive weeks. In patients exhibiting stable or regressive disease upon combined IL-2 and rIFN-, the therapy was continued. The overall response rate was 31% (95% confidence limits = 15%–51%), with 6 out of 29 patients achieving partial remission (PR, 21%) and 3 patients complete remission (CR, 10%). In addition, 12 patients presented with stable disease. The median duration of response was 8.5 months in PR and 19+ months in CR. Long-term treatment using this regimen was associated mainly with moderate (WHO grade I–II) toxicity including fevers, chills, malaise, nausea and/or vomiting, anorexia and transient local inflammation at the injection sites. No toxic deaths occurred. Altered thyroid function was observed in more than half the patients. The combination regimen resulted in a significant increase in peripheral blood eosinophils and natural killer cells (P<0.005). Up-on treatment, 14 patients developed non-neutralizing activity against rIL-2, and 2 of these developed specific neutralizing antibodies after consecutive cycles; no anti-rIFN-2b antibodies were detected. In summary, subcutaneous long-term outpatient treatment with low-dose rIL-2 and rIFN- is feasible, with moderate toxicity, and results in an objective tumor response rate comparable to that obtained previously with high-dose rIL-2 i.v. regimens.     

Sequential treatment of patients with advanced renal cell carcinoma with autologous tumor vaccine and subcutaneous administration of recombinant interleukin-2 and interferon α2b

Summary Thirty patients with advanced renal cell carcinoma (RCC), 23 of whom had distant metastases in at least one organ, were entered after nephrectomy into a protocol involving vaccination with Newcastle disease virus (NDV)-modified autologous tumour material, with a subsequent induction week and repetitive bi-weekly cycles of interleukin-2 (rIL-2) and interferon _2b/rIFN-_2b at a lower s. c. dose (1.5 million Cetus units m^–2 day^–1 every 12 h on 2 days and 3 million IU/m² once a day on days 1, 3 and 5). The inpatient treatment was followed by a maintenance phase during which 0.3 million Cetus units/m² rIL-2 was given s. c. every 12 h on days 1–5 and 3 million IU m^–2 day^–1 rIFN-_2b was administered on days 1, 3, and 5 on an outpatient basis. All but 3 patients completed the induction week and 6 weeks of outpatient therapy. No grade 3 or 4 toxicities occurred during the therapy. Therapy was discontinued for 3 patients because of rapid tumour progression. Of the 23 evaluable RCC patients, 3 exhibited a complete response and 4 displayed partial remission, 7 showed stable disease during 1–18 months (median = 5 months) of therapy, and progression was seen in 9. We conclude that vaccination with autologous tumour material combined with s. c. rIL-2 and rIFN-_2b administration can induce regressions in patients with advanced RCC and that even in non-responding patients a more favourable course of the disease can be achieved.     

Adoptive Immunotherapy With Tumor-Infiltrating Lymphocytes and Subcutaneous Recombinant Interleukin-2 Plus Interferon Alfa-2a for Melanoma Patients With Nonresectable Distant Disease: A Phase I/II Pilot Trial

Background: On the basis of our previous experience, we designed this study to determine the activity and toxicity of outpatient treatment with autologous tumor-infiltrating lymphocytes (TIL) together with intermediate-dose recombinant interleukin-2 (rIL-2) and low-dose recombinant interferon alfa-2a (rIFN-2a), for patients with metastatic melanoma.Methods: Between April 1992 and October 1994, we processed 38 melanoma samples derived from 36 patients with metastases. Proliferative cultures of expanded lymphocytes (TIL) were infused only once into patients with metastatic melanoma. rIL-2 was administered subcutaneously for 1 month, starting on the day of TIL infusion, at an escalating dose of 6–18 × 10⁶ IU/m²/day for the first week and at the maximum-tolerated dose for the subsequent 3 weeks and then, after a 15-day interval, for 1 week/month for 3 months. rIFN-2a was administered subcutaneously at 3 × 10⁶ IU three times each week until progression.Results: Of 38 melanoma samples, 19 (50%) resulted in proliferative cultures and were infused. The median number of expanded lymphocytes was 18 × 10⁹ (range, 1–43 × 10⁹), and the median period of culture was 52 days (range, 45–60). rIL-2 was administered at doses ranging between 6 and 18 × 10⁶ IU/m²/day. Toxicity was mild or moderate, and no life-threatening side effects were encountered. Two of 19 treated patients experienced complete responses of their metastatic sites (soft tissue), 10 had stable disease, and 7 showed progressive disease. The response rate was 11% (95% confidence interval, 2–35%).Conclusions: Outpatient treatment with TIL plus rIL-2 and rIFN-2a is feasible, although, within the context of the small sample size, the activity of the combination was no different from the reported activity of any of the components used alone.     

Phase II trial of interleukin 2, interferon α, and 5-fluorouracil in metastatic renal cell cancer: a cytokine working group study.

The purpose of this study was to evaluate the potential efficacy of alternating two outpatient regimens for the treatment of metastatic renal cell cancer. These regimens consisted of 4 weeks of recombinant interleukin 2 (rIL-2) plus IFN-alpha2B followed by 4 weeks of 5-fluorouracil plus IFN-alpha2B. Fifty patients meeting eligibility criteria of previous Cytokine Working Group studies were treated on an outpatient basis. Patients received s.c. rIL-2 (Proleukin; Chiron, Emeryville, CA) during weeks 1–4 of the 8-week regimen. During weeks 1 and 4, dosage for rIL-2 was 10 MIU/m² twice daily on days 3–5, and the dosage for IFN-alpha2B (Intron; Schering Plough, Kenilworth, NJ) was 6 MIU/m² on day 1. During weeks 2 and 3, the dosage for rIL-2 was 5 MIU/m2 on days 1, 3, and 5, and the dosage for IFN-alpha2B was 6 MIU/m² on days 1, 3, 5. During weeks 5–8, 5-fluorouracil (750 mg/m²) was administered once weekly by i.v. infusion, and IFN-alpha2B (9MIU/m²) was administered as a s.c. injection three times weekly. Throughout the treatment, an assessment of quality of life was made and a symptom-distress scale was evaluated. There were two patients with complete responses (CRs) and seven with partial responses (PRs) for an objective response rate of 18% (95% confidence interval, 10–25). The median response duration was 8 months (range, 3–51+ months). The CRs lasted 5 months and 51+ months and the PRs ranged from 3+ to 18 months. After completing at least one course of treatment, eight patients (three with PR, one with minor response, four with stable disease) became CRs after surgery for remaining metastatic disease. Six remain alive at 43+ to 53+ months, and 5 remain disease-free since surgery. The median survival of the study group is 17.5 months, with a maximal follow-up of 53+ months. The range in survival is 1–53+ months. Toxicity was primarily constitutional, and treatment modifications were designed to maintain toxicity at grade 2/3. The most toxicities during treatment with IL-2/IFN were fatigue, nausea/vomiting, anorexia, skin reaction, diarrhea, fever, and liver enzyme elevations. One-third had central nervous system toxicity (headache, depression, insomnia). During 5FU/IFN treatment, 49 of 50 patients experienced grade 2/3 myelosuppression during course 1. Eight patients experienced grade 4 toxicities. In conclusion, the activity of this alternating regimen is similar to that of IL-2/IFN alone, given in 4-week cycles. The addition of 5FU/IFN failed to increase the efficacy and added new toxicity (myelosuppression). This report does not confirm the results previously reported for either alternating or simultaneous administration of these three agents. Because 5FU does not appear to add to the antitumor activity of IL-2-based therapy for renal cancer, current efforts were directed toward a Phase III randomized comparison of high-dose i.v. bolus inpatient IL-2 treatment vs. treatment with outpatient s.c. injection of IL-2 plus IFN.     

Interleukin-2 and interferon-alpha2b as a daily alternating schedule in advanced renal cell cancer

Summary Interferon-alpha (IFN-) and interleukin-2 (Il-2) are effective as single agents in metastatic renal cell cancer (RCC) with response rates of 15–30%. Additionally, IFN- is assumed to act synergistically with Il-2 in the induction of lymphokine-activated killer cells. (LAK cells) in vitro. With the aims of increasing the response rate by combining both cytokines and of reducing side effects, we started a clinical trial with a daily alternating schedule of 10×10⁶ units/m² s.c. rIFN-_2b (Essex, Munich, FRG) and 3×10⁶ Cetus units/m² rIL-2 (EuroCetus, Frankfurt, FRG) in the form of 1 h infusions over a period of 14 days. Patients found to have progressive disease after two cycles of therapy were withdrawn from the study; patients with stable disease or better received two further cycles.Of the 27 patients included in the study, 22 (16 male, 6 female) are evaluable for response. In 1 patient with multiple pulmonary metastases complete remission was achieved, in 5 patients partial remission, and in 2 a minor response. The schedule was practicable; the main side effects were influenza-like symptoms, fatigue and hypotension. Some patients suffered from arthralgias and erythemas up to 3 weeks after finishing the therapy cycle. On the whole, the side effects seem to be less severe than those arising from schedules using continuous Il-2 infusions.This work was supported by Grant No. 01GA8802 of the Bundesministerium für Forschung und Technologie (BMFT)     

Chemoresistance of renal cell carcinoma: 1986–1994

Summary Multidrug resistance (MDR) in a variety of human tumors such as renal cell carcinoma (RCC) is thought to be caused by expression of the mdr1 gene and may be reversed by applying chemosensitizers such as Dexverapamil that inhibit the mdr1 gene product P-glycoprotein. On the basis of our preclinical analysis, we inititated a clinical (GCP) study with vinblastine (VBL), the most effective — if at all — chemotherapeutic agent; dexverapamil; and dexamethasone in patients with RCC. All patients had histologically proven RCC that was metastatic and progressive at study entry. The statistical design featured a preliminary study of two cycles of VBL alone followed by tumor evaluation. If no response was documented, with all patients thus serving as their own control, dexverapamil and dexamethasone were added for a minimum of three cycles of combination therapy. Having obtained institutional permission by the ethical review committee (MEC 124, 106-1993/12), we enrolled 24 patients on this protocol starting on May 3, 1993. In the preliminary study, 1 complete response (CR) was achieved with VBL alone, and myelotoxicity led to an adequate dose reduction from 2 mg/m² VBL per day given as a 5-day continuous infusion (days 1–5) in 6/10 yet evaluable patients to 1.4 mg/m² per day. In 8/11 yet evaluable patients, dexverapamil doses reached 3000 mg/day by 7-day oral uptake (days 0–6, supported by 20 mg dexamethasone given twice daily), which is significantly higher than those previously reported. The combination of VBL given at 1.4 mg/m² per day plus, dexverapamil given at 3000 mg per day was felt to be safe and well tolerated. Nine patients were yet evaluable for response. One partial response and three minor responses were noted in this heavily pretreated study population. It appears that this innovative approach may have some activity in RCC and may eventually lead to a rational treatment modality. Careful evaluation in ongoing studies is warranted.     

Human pancreatic growth hormone releasing factor (hpGRF): GRF-and GH-levels after bolus injection and infusion of hpGRF1–44

Summary Synthetic human pancreatic growth hormone releasing factor (hpGRF^1–44) was given as an i.v. bolus to healthy volunteers in 5 different dosages (3.3g to 200g hpGRF^1–44). In addition 11 healthy subjects were infused over 2 respectively 5 hours in a dosage of 100g hpGRF^1–44/h after receiving a bolus of 50 g hpGRF^1–44. Four healthy subjects served as placebo controls. GH, PRL, TSH, and GRF were measured by specific radioimmunoassays. The results show the clearcut dose response relationship between the administered GRF dosage and the resulting GH response from 3.3 to 50g hpGRF^1–44 i.v. Higher dosages of hpGRF^1–44 did not lead to a more pronounced GH response though there was a linear dose response relationship between the administered hpGRF^1–44 and the GRF immunoreactivity 5 minutes after injection. Infusion of hpGRF could not sustain elevated GRF levels and a second bolus of 50g hpGRF^1–44 given at the end of the 2-respectively 5-hour infusion led to a minor increase compared to the first bolus.100g hpGRF^1–44 was given to 14 patients with active acromegaly leading to a significant rise of the GH levels with the exception of 3 patients. Of the latter 3 two had received previous therapy, and one patient suffered from ectopic GRF hypersecretion. When GH responses to hpGRF^1–44 were compared to the responses to other releasing hormones there was no correlation. After transsphenoidal surgery divergent responses of GH were seen. In one patient with low basal GH and an exaggerated rise after GRF before surgery there was no response after successful transsphenoidal operation.We conclude that there is marked heterogeneity of the GH response to hpGRF^1–44 in normal subjects with a dose response relationship in the low dose range which is not apparent when higher dosages above 50 g hpGRF^1–44 are used. Long-term infusion can not sustain elevated GH levels which may be explained by antagonizing factors like somatostatin. Patients with active acromegaly who have not been treated show regularly a response to hpGRF^1–44, which may disappear after transsphenoidal surgery. In addition lack of GH responsiveness to hpGRF^1–44 does not exclude active acromegaly in already treated patients. If untreated patients do not have a GH response after hpGRF they may have an ectopic GRF source.     

Renal function and kidney size in glycogen storage disease type I

Renal failure has been reported recently as a late complication of glycogen storage disease type I (GSD I). We studied the renal function of 23 patients, mean age 10.9 years (range 2.2–21.6 years). The mean glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) were 188±50 and 927±292 ml/min per 1.73 m², respectively (normal values for adult controls 90–145 and 327–697, respectively). Hyperfiltration (GFR >145 ml/min per 1.73 m²) was found in 19 of 23 patients. There was no difference in GFR and ERPF between age groups 2–10 and 11–22 years. After a mean follow-up of 2.5 years (range 1–7.5 years) GFR and ERPF did not significantly change. At follow-up 3 patients (all older than 15 years) developed persistent glomerular proteinuria (0.1, 0.5 and 0.9 g/day). Besides a slight increase in fractional excretion of ₂-microglobulin (FE-₂m) in 6 patients, proximal tubular function tests (FE-₂m, tubular reabsorption of phosphate and glucosuria) were normal. In patients with increased kidney length related to body height, GFR and ERPF were significantly higher than in patients with normal kidney length. We conclude that GSD I is characterised by hyperfiltration and hyperperfusion. The relative increment in kidney length is related to the degree of hyperfiltration.     

Maturation of malfunctioning kidneys

Because loss of functional renal mass is compensated by hyperfiltration of remaining tissue, one could hypothesize that a damaged kidney might not have the same rate of maturation as the contralateral one. To verify this, maturation was evaluated in children with asymmetrical renal function during early life. Twenty-five children were selected having had 2 ^99mTc-MAG3 renograms combined with ⁵¹Cr-EDTA clearance measurement, enabling estimation of glomerular filtration rate (GFR), split renal function (SRF), and single kidney GFR (SKGFR). The first test had to be performed before the age of 18 months and SRF on the affected side had to be 40%. Moreover, GFR had to increase between the 2 tests by 10 mL^–1 min/1.73 m², reflecting maturation due to age. For 18 children SRF changed by between –5% and +5%. For 4 children an increase of 5% was observed whereas for the remaining 3 a decrease of 5% occurred. For the first 22 kidneys, mean increase of SKGFR was +6.3 mL^–1 min/1.73 m² (SD: 6). For the 3 kidneys with 5% SRF decrease, SKGFR remained unchanged in 2 (+0.3 and –3 mL min^–1/1.73 m²) and increased in the third patient (+15 mL min^–1/1.73 m²). In conclusion, renal maturation is comparable in both the malfunctioning kidney and the contralateral normal functioning side.     

Effects of calcium and temperature on tension in isolated canine coronary artery

The effects of calcium and temperature on the tension of isolated canine coronary arterial strips were studied.In 20mEq·l ^–1 K solution, the tension was significantly increased from 0mg with 0mEq·l ^–1 Ca to 33 ± 18mg with 0.2mEq·l ^–1 Ca at 37°C, from –40 ± 18mg with 0mEq·l ^–1 Ca to –17 ± 11mg with 0.2mEq·l ^–1 Ca at 30°C, from –77 ± 19mg with 0mEq·l ^–1 Ca to –52 ± 17mEq·l ^–1 with 1mEq·l ^–1 Ca at 25°C, from –88 ± 13mg with 0mEq·l ^–1 Ca to –41 ± 18mg with 2mEq·l ^–1 Ca at 20°C, from –125 ± 16mg with 0mEq·l ^–1 Ca to –116 ± 13mg with 2mEq·l ^–1 Ca at 15°C. Ca higher than 0.2mEq·l ^–1 produced a dose-dependent increase in tension between 37°C and 15°C. In spite of the presence of 4mEq·l ^–1 Ca, the development of tension was strongly supressed by lowering the temperature below 20°C, and completely inhibited at 10°C. The rate of a decrease in tension caused by cooling was about 5.5mg·°C^–1.This study demonstrated that Ca²⁺ produced a dose-dependent increase in tension in high-K solution, which was suppressed as the temperature was lowered.(Yoshida K, Fujii Y, Ina H, et al.: Effects of calcium and temperature on tension in isolated canine coronary artery. J Anesth 5: 172–176, 1991)     

Age-dependent alterations in the response of isolated rat aortas to Thiobarbiturates

Responses to thiamylal and thiopental were compared in helical strips of rat thoracic aortas of different ages (5–6, 10–12 and 20–22 weeks old), precontracted partially with phenylephrine or prostaglandin F₂ (PGF₂). Thiamylal and thiopental, in concentrations of 3 × 10^–5 to 10^–4M, produced a dose-dependent relaxation in aortas at 5–6 weeks of age, no significant change of tension in those at 10–12 weeks of age, and a marked constriction in those at 20–22 weeks of age. These thiobarbiturates, in a high concentration of 10^–3M, produced a profound relaxation in aortas at any age studied. It is concluded that the responses to thiobarbiturates of thoracic aorta precontracted with phenylephrine or PGF₂ alter with age.(Beak W, Hatano Y, Nakamura K, et al.: Age-dependent alterations in the response of isolated rat aortas to thiobarbiturates. J Anesth 5: 199–202, 1991)     

A bone resorbing substance from bovine serum albumin (brA)

Summary A fraction (brA), which causes resorption of fetal rat bones in vitro, has been concentrated from bovine serum albumin by anion exchange column chromatography on DEAE Sephadex. This active fraction has also been prepared using DEAE Sephadex A-50 by a batch method with a 0.09M NaCl, 0.1M TRIS buffer, pH 8.35. BrA was 10–30 times more potent than the original albumin. The retained material, which constitutes the bulk of the protein and has less activity than the original albumin, elutes with 0.45M NaCl. Similar treatment of serum, or globulins does not yield brA. Further enhancement of the bone resorbing activity of brA can be obtained with (NH₄)₂SO₄ fractionation or extraction with CH₃OHCHCl₃. Heating at 55° C for 2 h or at 100° C for 10 min does not affect the activity; overnight incubation with protease destroys the bone resorbing effect. The bone resorbing activity is not removed by dialysis and does not correlate with the protease activity of the fraction. The action of brA is inhibited by 3 mM PO₄, 1 g/ml calcitonin or glucagon, 10^–7 M dexamethasone or 0.02 g/ml actinomycin D. The bone resorbing activity of brA is partially inhibited by 10^–7–10^–5 M indomethacin. PTH did not elicit bone resorption when added to cultures incubated in chemically defined medium supplemented with 0.1 mg/ml brA. However, brA did not inhibit PTH-induced resorption.     

The effects of nicardipine given after 10-minutes complete global cerebral ischemia on neurologic recovery in dogs

The effect of nicardipine (NC) on neurologic recovery from ischemic insult after 10-minutes complete global cerebral ischemia was evaluated in dogs by examination of neurologic recovery score (NRS: complete recovery = 100, death = 0). Ischemia was achieved by occlusion of ascending aorta, and NC, 10µg·kg^–1 in bolus followed by infusion of 0.33µg·kg^–1·min^–1 for 2 hours, was administered immediately after re-establishment of circulation. The mortality at 7th day was 2/9 in the control © and 1/9 in the NC group (ns). NRS on 2nd day was 52.3 ± 6.8 in the C and 70.6 ± 6.5 in the NC (P 0.05), but that on 7th day did not differ between the two groups. The numbers of dogs recovered to over 80 in NRS on the 2nd day was 1/9 in the C and 5/9 in the NC (P 0.05), but that on the 7th day increased to 3/9 in the C and remained at 5/9 in the NC (ns). These results suggest that NC accelerates the early neurologic recovery from ischemic damage, but influences little the final outcome.(Iwatsuki N, Ono K, Takahashi M et al.: The effects of Nicardipine given after 10-minutes complete global cerebral ischemia on neurologic recovery in dogs. J Anesth 4: 337–342, 1990)     

Effect of tumor necrosis factor-α and interferon-γ on the growth of human prostate cancer cell lines

Human recombinant tumor necrosis factor- (rTNF-, 10^-12–10^-8 M) inhibited the proliferation of androgen-dependent LNCaP cells by 32–56%. In contrast, proliferation of androgen-independent PC-3 and JCA-1 cells was only slightly inhibited, or not inhibited at all, respectively. Human recombinant interferon- (rIFN-, 500 U/ml) decreased proliferation of PC-3 and JCA-1 cells by 35% and 53%, respectively, but had no effect on LNCaP cells. Interestingly, the combination of rIFN- and TNF- had greater antiproliferative effects on JCA-1 cells than treatment with either cytokine alone. However, the antiproliferative effects of this combination were similar to those observed for PC-3 or LNCaP cells treated with rIFN- or TNF- alone, respectively. These data suggest that some forms of androgen-independent prostate cancer may benefit from a combination therapy of IFN- and TNF-, while the use of IFN- alone may be more efficacious in others.     

Combined treatment of pelvic exenterative surgery and intra-operative pelvic hyperthermochemotherapy for locally advanced rectosigmoid cancer: Report of a case

A huge rectosigmoidal cancer which extended into the urinary bladder in a 64-year-old man is herein described. The tumor occupied the pelvic and lower abdominal cavities, while the rectosigmoid was totally obstructed. No hepatic or pulmonary metastasis was evident. The ventral and flank sides of the peritoneum in the right lower abdomen, right common iliac vessels, bilateral ureters, terminal ileum, cecum, ascending colon, and urinary bladder were all directly invaded by the tumor, but the aorta, sacrum, and lower rectum were free of cancer. Consequently, an anterior pelvic exenteration was carried out along with an ileal conduit and a right hemicolectomy. Immediately after the exenteration, intra-pelvic hyperthermochemotherapy was performed using a 46–47°C perfusate containing 40 g/ml of mitomycin C (MMC) and 200 g/ml of cisplatin (CDDP), for 90 min, in an attempt to prevent any further local recurrence. A right hemicolectomy and a permanent colostomy were done simultaneously with the hyperthermia treatment. After an uneventful postoperative course, the patient was prescribed adjuvant chemotherapy, i.e., two administrations of 17 mg/m² and 21 mg/m² of MMC, and ten doses of 710 mg/m² of 5-fluorouracil (5-FU) followed by five doses of 535 mg/m² of 5-FU. At the time of this writing, the patient is still alive without recurrence at 21 months after surgery.     

Inhibition of neutrophil superoxide production by adenosine released from vascular endothelial cells

To investigate the inhibitory effect of adenosine released by endothelium on neutrophil Superoxide (O ₂ ^–) production, we treated confluent monolayers of cultured human umbilical vein endothelial cells with the enzyme adenosine deaminase, and then added human neutrophils. Superoxide (O₂ ^–) production by human neutrophils stimulated with 10^–6 M formyl-methionyl-leucyl-phenylalanine was inhibited by 49% in the presence of a confluent monolayer of human umbilical vein endothelial cells (5.1 ± 0.1 versus 2.6 ± 0.3 nmols O₂ ^–/10⁶ neutrophils). Addition of 0.25 U/ml adenosine deaminase to neutrophils plus endothelial cells restored formyl-methionyl-leucyl-phenylalanine-stimulated neutrophil Superoxide production to the level seen with neutrophils alone. Deoxycoformycin (10^–4 M), an inhibitor of adenosine deaminase activity, prevented the increase in Superoxide production associated with adenosine deaminase addition. The adenosine analogue 5-(N-ethylcarboxamido)-adenosine (3 × 10^–4 M) caused increased inhibition of formyl-methionyl-leucylphenylalanine-stimulated superoxide release by neutrophils in the presence of endothelial cells and prevented neutrophil-mediated endothelial cell damage, as measured by release of³H-2-deoxy-D-glucose. Pairing 2-chloroadenosine (10^–5 M) or 5-(N-ethylcarboxamido)-adenosine (3 × 10^–4 M) with a cyclic adenosine monophosphate phosphodiesterase inhibitor, 3-isobutyl-l-methyl-xanthine (10^–4 M), produced greater inhibition of neutrophil superoxide production than occurred with either compound alone. The results support the hypothesis that vascular endothelial cells protect themselves from neutrophil attack by releasing adenosine to inhibit superoxide production.     

Interstitial photodynamic therapy in the Dunning R3327-AT6 prostatic carcinoma

Interstitial photodynamic therapy (PDT) could be an alternative radical treatment for prostate cancer. The ability to predict the depth of necrosis is necessary for light treatment planning using multiple optical fibres. The extent of PDT necrosis was studied in subcutaneously implanted R3327-AT6 Dunning prostate tumours which had similar optical characteristics to human prostate. Tumour-bearing subjects were given 20 mg kg^–1 Haematoporphyrin esters (HPE) and irradiated 24 h later with 630 nm laser light. Five subjects per group were treated with increasing light doses (50–450 J cm^–1) delivered interstitially via a single 2 cm long cylindrical diffuser. After 450 J cm^–1 of irradiation, 4.3±0.8 cm³ [standard error of the mean (s.e.m.)] of tumour tissue was necrosed to a depth of 10.5±0.8 mm around the diffuser. There was an approximately linear correlation between the volume of PDT necrosis around the fibre and prescribed light dose. The mean threshold light dose for PDT effect was 18±2 J cm^–2. In this tumour with a mean photosensitizer concentration of 16±1.5g g^–1, low light doses produced tumour necrosis. PDT using multiple diffusers could destroy a relatively large tumour volume and the diffusion theory model reliably predicted the depth of necrosis.     

Urinary endothelin excretion in the neonate: influence of maturity and perinatal pathology

The present study was undertaken to establish the developmental pattern of urinary endothelin-1 (ET-1) excretion and to define its possible role in mediating pathophysiological changes related to perinatal asphyxia/infection and dopamine treatment. Urinary ET-1 levels were measured by radioimmunoassay in 7 full-term neonates (mean gestational age 39.3 weeks) on days 1, 3 and 5, and in 9 pre-term neonates (mean gestational age 30.8 weeks) on days 1, 3, 5, 7 and weekly thereafter for 5 consecutive, weeks. The results were compared with those of three age-groups of 30 normal children (4–8 years, 9–12 years and 13–18 years); each group, consisted of 10 children. The influence of severe cardiopulmonary distress (n=16, mean gestational age 33.9 weeks, post-natal age 3.3 days) and dopamine administration in a dose of 2 g/min per kg (n=10, mean gestational and post-natal ages 32.1 weeks and 5.6 days, respectively) were also studied. In full-term infants, ET-1 concentration fell from 34.3±1.8 pmol/l on day 1 to 21.5±1.5 pmol/l on day 5 (P<0.01). In premature infants its absolute value and its post-natal fall were similar in the 1st week and no further change occurred in weeks 2–5; it stabilized at levels between 17.1±2.2 and 16.7±1.7 pmol/l. These concentrations tended to be lower than those of 25.5±1.3, 23.0±1.0 and 26.2±0.7 pmol/l measured in three groups of older children. During the 1st week, daily ET-1 excretion remained unchanged in term infants (3.1±1.0 vs. 3.7±1.5 pmol/m² per day), but there was a significant increase from 6.5±1.0 to 12.4±0.7 pmol/m² per day (P<0.01) in premature infants. During weeks 2–5, preterm infants excreted more ET-1 than older children (P<0.01). In response to perinatal ashphyxia/infection and dopamine therapy, urinary ET-1 excretion markedly rose and there was a significant positive correlation between urine flow rate and ET-1 excretion (P<0.001). We conclude that ET-1 concentration rather than excretion rate may have a role in mediating the changes in renal functions that occur soon after birth. The pathophysiological significance of the flow-dependent increase in urinary ET-1 excretion needs to be further studied.     

Alternating sequential chemotherapy with high-dose ifosfamide and doxorubicin/cyclophosphamide for adult non-small round cell soft tissue sarcomas

Doxorubicin and ifosfamide are the two most active agents used to treat soft tissue sarcomas. However, because of their overlapping side effects, concurrent administration to achieve optimal doses of each agent is difficult. We therefore conducted a Phase II trial to investigate the efficacy and feasibility of a novel alternating sequential chemotherapy regimen consisting of high dose ifosfamide and doxorubicin/cyclophosphamide in advanced adult non-small round cell soft tissue sarcomas. Adult patients with non-small round cell soft tissue sarcomas were enrolled. The treatment consisted of four sequential courses of chemotherapy that was planned for every 3 weeks. Cycles 1 and 3 consisted of ifosfamide (14g/m²), and cycles 2 and 4 consisted of doxorubicin (60mg/m²) and cyclophosphamide (1200mg/m²). Forty-two patients (median age 47 years) were enrolled. Of the 36 assessable patients, 1 complete response and 16 partial responses were observed, for a response rate of 47.2%. Responses were observed in 57% of patients who had received no previous chemotherapy and 13% of those who had previously undergone chemotherapy. Grade 3–4 neutropenia was observed during 70% of all cycles. Sequential administration of high-dose ifosfamide and doxorubicin/cyclophosphamide has promising activity with manageable side effects in patients with advanced adult non-small round cell soft tissue sarcomas.     

Changes in the plasma histamine concentration after the administration of vecuronium bromide

Clinical symptoms of anaphylactoid reaction to muscle relaxants vary from localized flush to cardiovascular collapse. Vecuronium bromide is reported to have very little histamine releasing property. However, there are some reports of anaphylaxis or anaphylactoid reaction to vecuronium. We studied plasma histamine concentration after the intravenous injection of vecuronium to confirm the histamine release. Twenty patients were randomly allocated to one of two groups, each group comprising of 10 patients: one group was to receive vecuronium 0.1mg·kg^–1 and the other 0.2mg·kg^–1 using the priming principle. Blood samples were taken prior to and 1, 3, 5, 8 and 13min after the administration of vecuronium. The plasma histamine concentration was measured by radioimmunoassay with monoclonal antibody.There were no significant changes in plasma histamine concentration over 13min after the administration of vecuronium compared with the baseline value. There were also no significant differences between these two groups. We concluded that vecuronium up to 0.2mg·kg^–1 did not change the plasma histamine concentration in the patients having no previous history of allergy or atopic tendencies.(Mitsuhata H, Matsumoto S, Enzan K, et al.: Changes in the plasma histamine concentration after the administration of vecronium bromide. J Anesth 5: 24–29, 1991)