Expression of the gene coding for the major outer capsid protein of SA-11 rotavirus in a baculovirus system.

The gene coding for the major outer capsid protein (VP7) of simian rotavirus SA-11 has been expressed in a baculovirus-insect cell system. The resulting protein is 35 kDa and is primarily associated with the endoplasmic reticulum. Neutralizing SA-11 polyclonal antiserum and VP7 monospecific antiserum reacted specifically with the expressed gene product. Antiserum derived against the recombinant VP7 protein neutralized SA-11 rotavirus infectivity in a fluorescent focus assay.

Clinical experience with immediate breast reconstruction using tissue expansion or transverse rectus abdominis musculocutaneous flaps.

We retrospectively reviewed our experience with immediate breast reconstruction in 103 consecutive patients with stage 0 or I breast carcinoma between May 1983 and April 1988. Two reconstructive techniques were used, that is, either tissue expansion with secondary prosthesis implantation (60%) or transverse rectus abdominis musculocutaneous (TRAM) flap (40%). Chemotherapy was administered in 22% of patients without delay or compromise. The mean length of follow-up is 30 months. The complication rate was equal for both groups (24%) with infection being most common in the group of patients with tissue expansion and partial flap necrosis being most common in the group of patients with TRAM flaps. Aesthetic results were superior with use of the TRAM flap. Our experience concurs with previous reports that documented satisfactory results with immediate breast reconstruction without compromising further therapy. We conclude that although the tissue expansion technique yields acceptable results, the TRAM flap yields superior aesthetic results in terms of both appearance and consistency.

Mammographic features of intracystic papillary lesions.

In this article, the mammographic findings of ten patients with intracystic papillary lesions are reported. Two of these patients also had sonograms, which confirmed the presence of intracystic papillary fronds. Although not all intracystic lesions can be differentiated from gross cysts, there are several helpful roentgenologic and clinical clues. The tumor presents mammographically as a sharply circumscribed cystic mass with an irregular and sometimes nodular contour, except where tumor breaks through the wall of the cyst to invade the parenchyma. There the borders become shaggy. This tumor should be suspected clinically if a cystic lesion is seen in a postmenopausal woman not taking estrogens. Management depends on a high degree of suspicion, and a sonogram is useful to visualize the papillary fronds in the cysts. Although malignant papillary lesions are often large on presentation, they carry an excellent prognosis that is not related to their size. Treatment consists of either mastectomy or lumpectomy and radiation, with or without dissection of axillary nodes.

Stage I and II breast carcinoma: treatment with limited surgery and radiation therapy versus mastectomy.

Between 1980 and 1986, 2,140 patients with surgical stage I or II breast carcinoma were treated including 1,179 patients with T1-2N0 disease and 961 patients with T1-2N1 disease. Among the 1,179 patients without node involvement, 215 underwent limited surgery (complete excision and axillary node dissection) and radiation therapy; 964 patients underwent modified radical mastectomy only. Of the 961 patients with node involvement, 106 were treated by means of limited surgery and radiation therapy; of these, 48 also received chemotherapy. The remaining 855 patients underwent mastectomy; of these, 381 also received chemotherapy and/or hormone therapy. The 5-year survival rates for patients with no node involvement were 96% for the group treated by means of limited surgery and radiation therapy and 88% for the group treated by means of mastectomy (P greater than .05). The 5-year survival rates for patients with node involvement were 96% for the group treated by means of limited surgery and radiation therapy with or without chemotherapy and 77% for the group treated by means of mastectomy with or without chemotherapy (P less than .01). This study demonstrates no disadvantage from treatment by means of limited surgery and radiation therapy and suggests that adjuvant radiation therapy may be important in increasing survival among patients with T1-2 breast carcinoma and positive axillary nodes.

Cloning and expression of the major inner capsid protein of SA-11 simian rotavirus in Escherichia coli.

The major inner capsid protein (VP6) of SA-11 simian rotavirus has been expressed in Escherichia coli using a cloned cDNA derived from SA-11 double-stranded RNA segment 6. The cloned gene was fused to the N-terminal coding sequence of lacZ resulting in the synthesis of a 44-kDa protein. Several smaller polypeptides were also observed, resulting predominantly from transcription and translation within the gene 6 coding sequence. The recombinant VP6 proved to be antigenic by immunoblot analysis using polyclonal serum against SA-11 rotavirus and by Western-blot analysis using monospecific serum derived from purified viral VP6.

Structure and organization of the gene encoding human pulmonary surfactant proteolipid SP-B.

Human pulmonary surfactant proteolipid SP-B arises by proteolytic processing of a 42,000-dalton precursor. The active proteolipid SP-B is one of two small hydrophobic proteins identified in surfactant that impart surface-active properties to surfactant phospholipids. We report the isolation and characterization of complete SP-B cDNA from a human lung cDNA library. The cDNA was used to isolate the gene encoding the SP-B precursor from a lambda EMBL3 library of human embryonic kidney DNA. The entire SP-B gene was sequenced and is approximately 9.5 kb long, with 11 exons and 10 introns including a large 823-nucleotide 3' untranslated exon. The sequence derived from the exons differs from the cDNA sequence at 3 positions out of 2001, only one of which is in the translated region. Direct RNA sequencing indicated that the 5' untranslated region is only 14 nucleotides long. A number of putative regulatory elements were found upstream of the SP-B gene, including a GC box and several putative cAMP and glucocorticoid receptor binding sites. Several Alu repeats and a region of potential Z-DNA formation were found in the introns. Southern blotting of human genomic DNA probed with SP-B cDNA indicated the presence of only one SP-B gene in the human genome, and the gene was localized to chromosome 2.

Two SP-C genes encoding human pulmonary surfactant proteolipid.

Human pulmonary surfactant proteolipid of Mr = 5,000, now termed surfactant protein C (SP-C), is produced by proteolytic processing of an Mr = 22,000 precursor. The active hydrophobic peptide imparts surface active properties to pulmonary surfactant phospholipids. We have determined the entire nucleotide sequence of two distinct genes encoding SP-C from a genomic library prepared from human leukocytes. SP-C genes were encoded by approximately 3.0 kilobase pairs of DNA containing six exons and five introns. In both genes, the active hydrophobic region of the polypeptide was located in the second exon that encodes a peptide of 53 amino acids. The entire nucleotide sequences of the two classes of SP-C genes differed by only 1%. Two cDNAs encoding SP-C were distinguished on the basis of an 18-nucleotide deletion at the beginning of the fifth exon; no such deletion was detected within the two classes of SP-C genes. Comparison of the 3'-untranslated regions of SP-C cDNA clones and the two classes of genomic clones demonstrated that cDNAs with and without the 18-base pair deletion could be derived from both of the genes. This 18-base pair deletion occurs in nucleotide sequences compatible with two distinct RNA splice sites. One additional cDNA clone showed the addition of an 8-base pair insert at the end of exon 5, which was also compatible with two distinct splice sites. Both classes of SP-C genes were represented by cDNAs, demonstrating that both classes of genes are actively transcribed. The two SP-C genes were readily distinguished on the basis of their nucleotide sequences and restriction fragment analyses of their flanking DNA. Two distinct classes of human SP-C genes are transcribed, and the heterogeneity in the SP-C RNAs appears to result from differential splicing.

Tumor to fascia margin as a factor in local recurrence after modified radical mastectomy.

A new problem has arisen for surgeons now that the pectoralis major muscle is routinely left in place after mastectomy. When the pathologist reports a tumor close to the fascial margin, there has been uncertainty regarding the significance of this finding. In the present study, the histories of 346 women with negative nodes who underwent modified radical mastectomy and had an uninvolved plane under the breast were reviewed. The distance from tumor to fascia was recorded by the pathologist, and the patients were divided into "Close" and "Not Close" groups. The "Close" group (90 patients) had tumors within one low power field (4 millimeters) of the fascia while the "Not Close" group (256 patients) had tumors more than 4 millimeters from the fascial margin. Twelve of the patients had local recurrence within an average follow-up period of 47 months, and a variety of analyses failed to show a statistically significant difference in local recurrence rates between the two groups. The results of this study indicate that tumor to fascia margin, as recorded by the pathologist, is not a strong determinant of local recurrence provided the areolar plane between the breast and the underlying fascia appears uninvolved at the time of mastectomy.

The extent and distribution of cancer in breasts with palpable primary tumors.

The term multicentricity has been employed to describe cancer cells beyond the borders of the primary tumor. However, it is not clear if there are multiple independent sites of origin or if the process simply represents spread of the cancer. The present study was designed to examine the distribution and extent of cancer in the breast and identify factors that bear on these events. All mastectomy specimens between 1980 and 1983 were systematically examined by means of multiple sections. One hundred seventy-nine of 657 patients (27%) were found to have separate foci. The most common histologic type (invasive ductal) was least likely to have multifocal disease (19%), while it was extremely common in the small group of patients with intraductal lesions (81%). Size was a factor in ductal but not in lobular lesions. Ninety per cent of the secondary foci were found in close proximity to the primary, suggesting spread rather than multicentricity. This implies a more limited and predictable distribution of cancer cells and opens the way to more rational selection and surgical preparation of patients for breast preservation.

Non-human primate (baboon) anti-gross cystic disease fluid protein-15 antibody infusion in four women with metastatic breast carcinoma.

Four women with metastatic breast carcinoma and elevated plasma levels of human breast gross cystic disease fluid protein of 15,000 dalton monomer size (GCDFP-15) were treated IV with non-human primate (baboon) anti-GCDFP-15 antibody. Three patients were given a single IV infusion of antibody, while the fourth patient received four sequential IV infusions. Antibody dosage patients, after antibody infusion the plasma level of GCDFP-15 decreased to 0 ng/ml and remained there as long as "free" circulating anti-GCDFP-15 antibody was present. The plasma half-life of the antibody ranged between 1 and 40 h and the duration of detectable free antibody ranged from 6 to 240 h. No toxicity was observed for the dosage range of antibody tested. No anti-baboon antibody response was detected. In the patient who received four sequential infusions of antibody partial regression of subcutaneous metastatic nodules occurred. The other three patients showed no clinically detectable changes from the antibody infusion.

The psychological impact of immediate breast reconstruction for women with early breast cancer.

Twenty-five patients were evaluated, 13 who had immediate breast reconstruction and 12 who had delayed breast reconstruction for early breast cancer. Data were elicited about the psychological impact of the cancer, the mastectomy, and the reconstruction. Our results support the conclusion that immediate breast reconstruction is accompanied by a lower incidence of psychological morbidity postoperatively, and we recommend that immediate breast reconstruction be offered as an alternative to women with early breast cancer.

Breast gross cystic disease fluid analysis. I. Isolation and radioimmunoassay for a major component protein.

Human breast gross cystic disease (GCD) fluid was analyzed by sodium dodecyl sulfate-acrylamide gel electrophoresis, and four major proteins (GCDFP-70), GCDFP-44, GCDFP-24, and GCDFP-15) were identified. By fractionation techniques, these proteins were separated from one another. The GCDFP-70 was immunologically identical to human albumin and was present in GCD fluid at approximately a 100-fold lower concentration than in plasma. The GCDFP-44 was immunologically identical to human plasma Zn-alpha2-glycoprotein; however, it was present in GCD fluid at an approximately 50-fold higher concentration than in plasma. The GCDFP-24 was the major component protein of GCD fluid. It had progesterone binding activity, and immunologically it was identical to a component of human plasma; however, antisera that identified 30 separate components of plasma failed to identify the GCDFP-24 as one of these plasma proteins. The GCDFP-24 concentration in GCD fluid was approximately 100-fold higher than the plasma analog. The GCDFP-15 component was immunologically distinct from any plasma components, as judged by Ouchterlony analysis. It was, however, immunologically identical with a component of both human milk and saliva. As revealed by radioimmunoassay, plasma levels in normal subjects were 7-85 ng/ml. In patients with metastatic breast carcinoma, markedly plasma levels (150-30,000 ng/ml) of this protein were detected. Short-term tissue cultures of breast carcinoma explants released this protein into the culture medium.

Evaluation of carcinoembryonic antigen as a plasma monitor for human breast carcinoma.

Plasma CEA levels have been determined in 92 normal women and 768 women with benign or malignant breast diseases. Only one of 92 normal women had a CEA level above 5 ng/ml. Of 253 women with benign breast diseases (gross cystic disease, adenofibroma, fibrosis, etc.) only one had a CEA level above 5 ng/ml. Ninety-four percent of the above two groups of women had CEA levels below 3 ng/ml. Of 164 women operated upon for Columbia Clinical Classification Stage A or B breast carcinoma, preoperative CEA levels were above 5 ng/ml in seven (4%). Patients with a preoperative CEA level above 3 ng/ml seemed to have an increased incidence of tumor recurrence. Elevated CEA levels (greater than 10 ng/ml) in our postmastectomy population of 288 patients have correlated with development of metastases in 14 of 46 subjects. Of 216 patients under treatment for metastatic breast carcinoma, CEA levels above 10 ng/ml have been detected in 15 percent of patients with soft tissue metastases, 38% of patients with visceral metastases and 50% of patients with osseous metastases. Of metastatic breast carcinoma patients with CEA levels above 10 ng/ml serial measurements have correlated with the patients response to therapy, progressively increasing in treatment failures and decreasing in treatment responders.

Comparative evaluation of carcinoembryonic antigen and gross cystic disease fluid protein as plasma markers for human breast carcinoma.

A comparative evaluation of carcinoembryonic antigen (CEA) and gross cystic disease fluid protein (GCDFP) as plasma markers for human breast carcinoma has been performed. Both assays appear to be useful in patients with metastatic breast carcinoma. Of 216 patients under treatment for metastatic disease, 111 (51%) had abnormal plasma levels of CEA and/or GCDFP. Abnormal plasma levels of CEA were present in 73 patients whereas abnormal GCDFP levels were present in 67. Twenty-nine patients had increased plasma levels of both markers simultaneously, 44 patients had increased CEA levels only and 38 patients had increased GCDFP levels only. Thus, of the 111 patients with elevated levels of either CEA or GCDFP, the two markers varied independently of each other in 74%. Utilizing both assays, abnormal plasma levels were present in 79% of patients with osseous metastasis, 53% of patients with visceral metastasis and 26% with soft tissue metastasis. Both assays, when performed serially in patients treated for metastatic breast carcinoma, were found to have utility in monitoring responsiveness; and increasing CEA or GCDFP plasma level indicated disease progression and a decreasing plasma level indicated regression.

Evaluation of a breast cyst fluid protein detectable in the plasma of breast carcinoma patients.

A radioimmunoassay has been developed for one of the major proteins isolated from human breast cyst fluid. Immunologically this protein is identical to a protein present in both human milk and saliva. Ninety-two normal women had plasma levels of this protein below 100 ng/ml (range 7-81 ng/ml; mean 31 ng/ml), and 85% had plasma levels below 50 ng/ml. More than one-half of the women with active gross cystic disease of the breast had plasma levels above 50 ng/ml. None of these women, however, had plasma levels above 150 ng/ml. A significant percentage of women with advanced breast carcinoma have been found to have plasma concentrations of this protein greater than 150 ng/ml.

In vitro stimulation of human breast tissue by human prolactin.

Normal human breast tissue was cultured with defined media plus hormones. The epithelium survived at least 4 days in culture but did not grow in the absence of hormones. Both insulin and human prolactin stimulated growth, but ovine prolactin did not.