Insecticidal proteins from Bacillus thuringiensis protect corn from corn rootworms.

Field tests of corn co-expressing two new delta-endotoxins from Bacillus thuringiensis (Bt) have demonstrated protection from root damage by western corn rootworm (Diabrotica virgifera virgifera LeConte). The level of protection exceeds that provided by chemical insecticides. In the bacterium, these proteins form crystals during the sporulation phase of the growth cycle, are encoded by a single operon, and have molecular masses of 14 kDa and 44 kDa. Corn rootworm larvae fed on corn roots expressing the proteins showed histopathological symptoms in the midgut epithelium.

Treatment of dermatomyositis with intravenous gammaglobulin.

PURPOSE: The mainstay of pharmacologic therapy in patients with dermatomyositis is corticosteroids. However, because patients sometimes become refractory to these drugs and because these drugs have potential short- and long-term toxicities, alternate therapy is highly desirable. Therefore, a pilot study was initiated using high-dose intravenous gammaglobulin (IVGG) in the treatment of dermatomyositis. PATIENTS AND METHODS: IVGG was administered to five patients with juvenile dermatomyositis. Prior to IVGG treatment, all patients had persistent muscle weakness despite daily corticosteroids and three patients had developed unacceptable steroid toxicity. Two of the patients had previously developed toxicity while receiving immunosuppressive therapy. RESULTS: IVGG therapy resulted in improved muscle strength and ameliorated skin rash in all patients. The percentage increase in muscle strength as measured by sphygmomanometry following the 9-month course of IVGG ranged from 56% to 606% in the proximal lower extremities and from 30% to 186% in the proximal upper extremities. Following IVGG therapy, prednisone could be discontinued or the dose reduced in all patients. CONCLUSION: This study suggests that IVGG may allow steroid sparing in dermatomyositis and may provide a safe alternative to cytotoxic therapy.

A non-competitive enzyme immunoassay of human alpha-lactalbumin in biological fluids and tissue extracts.

A sandwich non-competitive enzyme immunoassay for the measurement of human alpha-lactalbumin in serum, milk, and tumour tissue cytosol was developed using affinity-purified polyclonal antibody adsorbed to solid phase. The detection limits of this procedure in tubes (macromethod) and in wells of microtitre plates (micromethod) are 50 pg and 25 pg/sample, respectively, which means 2.5 micrograms/l of serum at appropriate dilution. The micromethod enables a reduction of the volumes used in the reaction with equal sensitivity, accuracy and reproducibility. A comparison of alpha-lactalbumin concentrations in serum of healthy, pregnant, and lactating women determined by our method with those measured formerly by radioimmunoassays proved the specificity of our procedure.

Longitudinal follow-up circulating immune complexes of the serum in patients with breast cancer.

The values of circulating immune complexes (CIC) in patients with breast cancer were followed for one year at 4-month intervals by means of the "PEGIKEM" test. From the total of 67 patients, 37 were treated surgically and a significant decrease in mean values of CIC occurred 8 months after the operation. 30 patients with a more advanced stage of the disease treated with radiotherapy and chemotherapy showed a decrease in mean CIC values already after 4 months but later on the CIC values increased again. No significant differences in the mean CIC values prior to treatment between individual clinical stages were found, only a certain tendency towards a more frequent occurrence of higher CIC values in more advanced stages was noted. Nor were the findings in lymph nodes reflected in the mean CIC values prior to treatment. Comparison of CIC values prior to treatment and the results of 2-hour blood sedimentation, revealed conformity in only 40% of the patients. In the other cases higher values of blood sedimentation were more frequent. The study of prognostic value of CIC after 3 and 5 years follow-up is in progress.

Haptoglobin type in connection with mean age at first breast cancer signs?

In breast cancer patients divided into groups according to haptoglobin (Hp) type (519 patients), blood group (ABO) (433 patients) or Rh factor (358 patients) the mean age at first disease manifestation was calculated. The results indicate that in Hp 1 bearers breast cancer appears at significantly lower age (49 years) as compared to Hp type 2-1 (53.3 years) or Hp 2 (53 years). In blood groups the mean age at first cancer signs raises from 48 years in the AB group to 53.1 years in the A group, only the difference between A and AB being significant. However, these preliminary results require confirmation in larger patient series.

Three-year follow-up of carcinoembryonal antigen levels in the serum of patients with breast cancer.

The levels of carcinoembryonal antigen (CEA) were followed up for three years in 66 patients with breast cancer. In the group of patients treated surgically (N = 36, T1-2, NO-2, MO) the values of CEA were in agreement with the clinical course in 97% of the patients (35 out of 36 patients). Recurrence and/or dissemination of the disease were signalled by a rise in the serum CEA level to more than 15 micrograms/1 out four out of five patients more than 3-10 months before clinical manifestation. In the group of patients without surgery, who were treated by radiation and/or chemotherapy (N = 30, T2-4, NO-3, MO) the serum CEA values were in agreement with the clinical course in 71% of the patients (20/28). Within three years recurrence and/or dissemination were detected in 57% patients (16/28), ten patients died (10/30--two of them were not evaluated). The recurrence or dissemination were signalled in seven out of 16 patients (44%) more than 4-14 months before clinical detection, in three patients (19%) the rise was detected simultaneously with the clinical finding (examinations at 3-4 months intervals), and in 6 patients (38%) the CEA level was at variance with the clinical finding.

Immunohistochemical localization of alpha-lactalbumin in human breast cancer tissue.

One hundred and eleven formalin-fixed breast cancer tissue samples were examined for the presence of alpha-lactalbumin using our polyclonal antibodies to this specific milk protein. Alpha-lactalbumin was shown to be present in 67 tumor samples (60%), in 15% of cases the occurrence of alpha-lactalbumin was questionable. No relationship was found between alpha-lactalbumin positivity and the histological type of the tumor, differentiation grading, type of invasivity, or stromal reactivity. Disease progression occurred in an equal number of patients with tumors either alpha-lactalbumin positive or negative. However, negativity was often connected with a disease-free interval shorter than 24 months after primary operation.

Occurrence of carcinoembryonic antigen in tumor tissue and serum of breast cancer patients.

In 116 breast cancer patients, the levels of carcinoembryonic antigen (CEA) were determined before operation in the serum using RIA, and after operation in sections of breast tumor tissue using the immunohistological PAP technique. CEA circulating in the serum was found in 49 patients (42%). Elevated values (over 10 micrograms/l) were found in only 12 patients (10%). In histological specimens CEA positivity was found in 94 tumors (81%), however, in a majority of them the number of positive cells per section was low (1-10%). A comparison of positive and negative findings both in the serum and in the tumor specimens of individual patients showed that both serum and tumor sections were CEA positive in 40 patients (35%) and both localizations were CEA negative in 13 patients (12%). Although most patients had positive histological sections but negative sera (46%). Only 7% of patients had negative sections and positive sera. In 41 patients CEA could be examined both qualitatively (immunohistologically), and quantitatively in the cytosol of the same homogenized tumor. Of them, 30 patients (72%) had in the cytosol a CEA concentration exceeding 5 micrograms/g proteins, in 11 of the 41 patients (28%) no CEA was found. Immunohistological examination of CEA in this group gave positive results in 35 out of the 41 patients (85%), and only 6 tumors (15%) were completely negative. CEA was shown to be present in each histological type of the tumors studied, invasive ductal tumors being slightly more frequent and more positive than the lobular ones. No relation was observed to the structure of the tumors, nor to the degree of their differentiation. Thus, the examination of CEA levels can hardly contribute to the improvement of histological classification.

Phylloid breast tumors and three steroid hormone receptors.

The concentrations of three steroid hormones (estrogen, progesteron and 1,25-dihydroxycholecalciferol) receptors (ER, PgR, DR) in tissue cytosol were analyzed in a group of 17 breast phylloid tumors. Comparison with breast carcinoma tissue samples (n = 37) did not reveal significant differences in average values of ER, PgR, and DR. Comparison with another control set of 30 samples of dysplastic tissue of the mammary gland showed significant differences only in PgR values. Only 18% of phylloid tumor samples contained levels above cut-of-line of all three receptors (ER, PgR, DR-5,10,10 resp. fmol/ mg protein). The most frequent combination was ER+PgR+DR-(41%). As far as we know, DR in phylloid breast tumors have never been examined before. In approximately 60% of our samples we found the expression of DR, in 36% the estimated values were above 10 fmol/mg protein. Cells of the tissue not expressing DR seem to belong to a special phenotype. We found no ER+PgR- or ER-PgR-combinations in them. The group which expresses DR is characterized by a higher dispersion of PgR values.

Carcinoembryonic antigen and alpha-lactalbumin in phylloid tumor tissues.

The concentration of carcinoembryonic antigen and alpha-lactalbumin in tumor tissue cytosol were analyzed in a group of 19 tumors of cystosarcoma phyllodes type. Both antigens were also localized in the tissue of identical tumors by means of immunohistochemical procedure. The cytosol levels of both proteins were found to be higher in the histologically defined malignant type of phylloid tumors. This group was also characterized by the simultaneous occurrence of both antigens. We did not manage to prove any relationship between the presence of alpha-lactalbumin and the steroid hormone receptor positivity in tumor tissue.

Complex biochemical analysis of human breast tumor tissue.

The quantitative biochemical analysis of tissue specimens from 76 human breast carcinomas consisted of examination for cytosolic estrogen receptors (cER), nuclear estrogen receptors (nER), progesterone receptors (PgR), 1,25-dihydroxycholecalciferol receptors (DR), carcinoembryonic antigen (CEA), alpha-lactalbumin (aLA), and gamma-glutamyl transferase (gGT). The highest incidence was found in CEA (76%), DR (70%), and aLA (62%). There was a high percentage of tumors containing only DR, in contrast to the tumors containing only cER or PgR. The simultaneous occurrence of DR and CEA was considerably high (61%). No statistically significant differences were observed in these biochemical parameters in relation to the grade of differentiation of the tumors. The values of aLA in tumors that invaded lymphatic or blood vessels were lower as compared to those tumors that invaded adipose or connective tissues. The level of statistical significance of this difference was close to 5%, the differences in other parameters were statistically insignificant. For prognosis assessed at the time of surgery, after a 2-3-year follow-up of 36 patients the level of gGT in the tumor seems to be the most promising prognostic factor. The values of gGT were significantly lower in those patients whose tumors were in progression during this time. The significance of nER and aLA was also taken into consideration.

The influence of certain colchicine derivatives on the growth characteristics of L cells.

Colchicine and its derivatives (demecolcine, 2-demethyldemecolcine and acetylated 2-demethyldemecolcine) blocked the mitotic activity of L cells and inhibited proliferation and DNA synthesis. This effect depended on the length of the exposure and the concentration of the substances tested (from 0.05 micrograms/ml up to 2.6 micrograms/ml). The greatest values of plating efficiency, cell proliferation, mitotic index and inhibition of DNA synthesis, respectively, were found after the application of colchiciline (0.2 micrograms/ml). These effects were reversible, since L cells underwent regeneration of the mitotic spindle and renewal of their function after the removal of colchiciline. Reversible effects were much less with other colchicine derivatives tested. The different effects of colchicine and its derivatives were due to their chemical structure, which determines the interactions with respective L cell receptors and thus modifies the mechanism of action on the mitotic apparatus of L cells.