Three forms of monocyte-derived neutrophil chemotactic factor (MDNCF) distinguished by different lengths of the amino-terminal sequence

Human monocyte-derived neutrophil chemotactic factor (MDNCF) was purified from culture supernatant of lipopolysaccharide-stimulated human peripheral blood mononuclear leukocytes on a column of Sepharose-bound murine monoclonal anti-MDNCF. About 65% of the culture fluid chemotactic activity was bound to the column. The unbound 35% probably represents chemotactic activity of other cytokines in the culture fluid. More than 85% of the bound activity was eluted by pH 2.5 glycine buffer. When this material was applied to an HPLC-CM column, gradient elution produced four well-separated A280 peaks, each of which had chemotactic activity. N-terminal amino acid analysis of the four peaks revealed three different sequences. One (MDNCF-c) was identical to the sequence that we reported previously. The other two (MDNCF-a and -b) had seven and five additional amino acids, respectively, at the N-terminus. MDNCF-a, -b and -c accounted for 8, 47 and 45% of the total MDNCF peptide. Alignment with the MDNCF cDNA sequence shows that MDNCF-a results from cleavage of a 20 residue signal peptide. MDNCF-c results from culture fluid proteolytic cleavage of the N-terminal sequences of MDNCF-a and -b at an R-S bond. The three peptides occurred in the four HPLC-CM peaks in different ratios. The bulk of any one peptide was distributed in two adjacent HPLC-CM peaks. This suggests that each peptide exists in a minimum of two states. In contrast to our previous multi-step purification, the immunoaffinity and HPLC-CM column sequence resulted in complete purification of MDNCF in two steps and led to identification of two additional MDNCF peptides, one of which has not heretofore been detected.     

Separation of human basophils into two fractions with different density and histamine content

We designed experiments in this study to test the hypothesis suggested by recent purification data that blood basophils comprise two populations of different density, which circulate in numbers characteristic for each human subject. Basophils were separated into two density bands by single step centrifugation on a discontinuous Percoll gradient. Band 1 cells were at the interface between plasma and Percoll of density 1.070 gm/ml. Band 2 cells were at the Percoll 1.070 to 1.080 interface. When the number of band 1 basophils was expressed as a percentage of the total in bands 1 and 2, this relative amount generally remained in a narrow range for blood obtained from the same donor on 3 successive days but differed markedly in different individuals. In a series of leukapheresis experiments, we demonstrated that the percentage of band 1 basophils in postleukapheresis venous blood was strikingly similar to the preleukapheresis value. If basophils that repopulated the leukapheresis-depleted circulation came from the bone marrow, we can conclude that blood levels of basophils in bands 1 and 2 are under physiologic control and that the two types of basophils are released in amounts characteristic for each human subject. Additional evidence for two distinct blood basophil populations was provided by histamine measurements. The histamine content per basophil was consistently higher in cells from band 1 than from band 2, the mean difference between pairs of values for 30 subjects being 0.3 +/- 0.04 pg or about 27% of the band 1 basophil histamine content of 1.1 pg.     

Radiation therapy and 5-fluorouracil modulated by leucovorin for adenocarcinoma of the pancreas. A phase I study.

This Phase I study was designed to build on the Gastrointestinal Tumor Study Group's experience with combined modality therapy in patients with pancreatic cancer. Thirteen patients with adenocarcinoma of the pancreas received weekly 5-fluorouracil by rapid intravenous infusion midway through a 2-h infusion of high dose leucovorin during external beam radiation therapy. Twelve patients received 100% of planned external beam radiation; treatment delays occurred in only three. Four patients received 100% of planned chemotherapy doses. Leukopenia and thrombocytopenia caused reduction of the number of chemotherapy doses given during radiation in six patients; diarrhea, severe nausea and vomiting, and wound abscess caused reduction in three patients. Ten patients were evaluable for response; two had complete responses, one had a partial response, and two had minor responses. In this small series baseline and post-treatment CA 19-9 levels predicted and correlated with response. We conclude that radiation and 5-FU modulated by leucovorin is a tolerable treatment regimen for carcinoma of the pancreas, with preliminary suggestion of activity, that warrants further Phase II testing.     

A phase II study of carboplatin and CHIP in patients with metastatic colon carcinoma

Fifty-six patients were treated in each arm of a study comparing CHIP and carboplatin for the therapy of previously untreated metastatic colorectal carcinoma. There were one partial response (2%) with CHIP and two partial responses (4%) with carboplatin. Side effects were significantly more severe with CHIP than with carboplatin. The most common side effect for both drugs was vomiting followed by hematologic side effects. Sixteen percent of the patients receiving CHIP and 9% of those receiving carboplatin had life-threatening side effects. Neither drug offers significant activity in metastatic colorectal carcinoma.     

A high molecular weight chemoattractant generated from C5 by ultracentrifugation of mouse serum without activation of complement

After ultracentrifugation of normal mouse serum, we found chemoattractant activity in the high molecular weight protein region at the bottom of the tube. which was comparable in amount and potency to the attractant in endotoxin-activated serum. This was not a pre-formed attractant, but was generated from serum reactants at least one of which was inactivated by heating at 56 C. Analysis of sera from 10 different mouse strains for hemolytic C5 activity and for capacity to generate chemoattractant on ultracentrifugation showed that the 4 strains without C5 were the only strains that failed to generate the attractant. Thus. the attractant precursor is C5. Since the activity was generated in the presence of 0.01 M EDTA, classical or alternative complement activation was not required. The chemoattractant product had a mol. wt of approximately 170,000; it was therefore not free C5a. These results, and data recently published on digestion of purified human C5 by trypsin. suggest that limited proteolysis of C5 can produce a chemoattractant molecule without release of free C5a.     

72 kidney transplants at the Medical College of Georgia 1968-1976.

Seventy-two kidneys have been transplanted-47 from related donors, and 25 from cadaver donors-into 63 recipients. No donor died or experienced a complication requiring more than one extra week of hospitalization. Six recipients died. Three of the six recipients died from complications. Two of the six died only because they declined to resume hemodialysis, and one died in prison of unusual circumstances after seven and one-half years of good health and kidney function. Five patients are surviving by virtue of a second transplant. Eight patients are now on hemodialysis. Seven patients with transplants are not in excellent physical health and five other patients with excellent health and renal function are nonetheless frequently depressed and easily upset over minor physical disabilities. A majority of the recipients have been rehabilitated to a gratifying extent.     

A phase II study of bisantrene in advanced refractory breast cancer. An Eastern Cooperative Oncology Group pilot study

Thirty patients with advanced refractory breast cancer received bisantrene 260 mg/m2 intravenously every 3 weeks. Reversible myelosuppression was the most commonly observed side effect. Four patients (13.3%) achieved objective partial response (90% confidence intervals 3-24%), while two patients (6.6%) had disease improvement with a PR + IMP rate of 19.9%. Seven additional patients (23.3%) had stabilization of disease. This drug has antitumor activity against breast cancer and warrants further study, particularly if problems with drug delivery are overcome.     

Understanding locally advanced breast cancer: what influences a woman's decision to delay treatment?

BACKGROUND: Despite early detection having been identified as a key factor in long-term survival of patients with breast cancer, women with locally advanced breast cancer continued to present at many medical centers. METHODS: Women with locally advanced breast cancer (LABC) (N = 11) and matched controls (N = 11) with Stage I or II breast cancer completed semi-structured interviews and the Hospital Anxiety and Depression Scale, Life Orientation Test, Multidimensional Health Locus of Control Scale, Body Investment Scale, Monitor-Blunter Style Scale, and Religious Coping Scale. Five spouses of the LABC group and eight control partners were similarly assessed. RESULTS: On psychological tests, no significant differences were found between patient groups; however, LABC spouses reported significantly greater active religious surrender (t = 2.37, P = 0.037) and depression (t = 3.54, P = 0.047) than control spouses. The women's semi-structured interviews identified that LABC was associated with inattention to routine breast cancer screening, denial, fatalism and reliance on alternative therapies. Spouses of the LABC group tended to be more passive in their wives' medical care, and also utilized fatalistic thinking and denial. CONCLUSIONS: It is important for health care providers to identify patients who will require extra encouragement to seek timely medical care and to follow through with treatment recommendations.