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PubMed2006

Selection and characterization of Her2 binding-designed ankyrin repeat proteins.

Designed ankyrin repeat proteins (DARPins) are a novel class of binding proteins that bind their target protein with high affinity and specificity and have very favorable expression and stability properties. We describe here the in vitro selection of DARPins against human epidermal growth factor receptor 2 (Her2), an important target for cancer therapy and diagnosis. Several DARPins bind to the same epitope as trastuzumab (Herceptin), but none were selected that bind to the epitope of pertuzumab (Omnitarg). Some of the selected DARPins bind with low nanomolar affinity (Kd=7.3 nm) to the target. Further analysis revealed that all DARPins are highly specific and do not cross-react with epidermal growth factor receptor I (EGFR1) or any other investigated protein. The selected DARPins specifically bind to strongly Her2-overexpressing cell lines such as SKBR-3 but also recognize small amounts of Her2 on weakly expressing cell lines such as MCF-7. Furthermore, the DARPins also lead to a highly specific and strong staining of plasma membranes of paraffinated sections of human mamma-carcinoma tissue. Thus, the selected DARPins might be used for the development of diagnostic tests for the status of Her2 overexpression in different adenocarcinomas, and they may be further evaluated for their potential in targeted therapy since their favorable expression properties make the construction of fusion proteins very convenient.

Amino Acid Sequence↗
PubMed1993

Effects of high-fat diet on incidence of spontaneous tumors in Wistar rats.

In a 2.5-year carcinogenicity study, two groups, both including male and female Wistar rats, were fed two different diets with 4% and 16% fat. In addition to 4% soybean oil, the high-fat diet contained 12% mono- and diglycerides, of which 85% was stearic acid and 13% palmitic acid. There was no difference in food consumption, body weight, weight gain, and longevity between the two groups. A statistically significant increase in the incidence of tumors in the high-fat group was seen in fibroadenoma of the mammae (female, p = 0.05). No statistically significant difference was seen when the incidence of benign mammary tumors (adenomas and fibroadenomas) was combined, just as the overall incidence of mammary tumors (adenomas, fibroadenomas, and adenocarcinomas) was not significantly different between the groups. A statistically significant decrease in the incidence of tumors in the high-fat group was seen in adenoma of the parathyroid gland (male, p = 0.04) and medullary carcinoma of the adrenal gland (male, p = 0.04). Combining the incidence of benign and malignant tumors of the adrenal medulla led to a further increase in the level of significance (p = 0.02). The present study showed that a high-fat diet influenced the tumor incidence in certain organs of rats. However, the overall differences in tumor incidence between rats fed the low- and the high-fat diet are considered marginal. Therefore we were not able to confirm or deny the hypothesis that a high-fat diet promotes the development of cancer. It should be noted that, in our study, fat accounted for about 30% of the total energy in the high-fat diet. This is much below the amount of fat normally found in the western diet but corresponds well to the level recommended for human intake. In addition, the rats fed the high-fat diet did not gain more weight, even though no difference was recorded in food consumption (g/kg body wt) between the groups.

Adenofibroma↗
PubMed1995

Refractoriness to mammary tumorigenesis in parous rats: is it caused by persistent changes in the hormonal environment or permanent biochemical alterations in the mammary epithelia?

Administration of a single i.v. injection of 50 mg N-methyl-N-nitrosourea (MNU)/kg body wt to 50- to 60-day old virgin rats, 120-day-old virgin rats, and 120-day-old parous rats (Sprague-Dawley; n = 18-37) resulted in a high incidence of mammary carcinomas in the virgin animals (97.3% in 50- to 60-day-old virgin rats; 75.0% in 120-day-old virgin rats), but mammary carcinomas did not develop in the parous rats. The concentrations in serum of various mammotropic hormones were measured in identical groups of rats at the time of MNU treatment. Growth hormone (GH) concentration was significantly reduced in parous rats, as compared with young or age-matched virgin rats. The concentrations of prolactin, 17 beta-estradiol, progesterone, corticosterone and thyroxine were not significantly altered in the parous rats compared to the two groups of virgin animals. Histological examination of the mammary glands from the three groups of rats showed that the epithelia of the parous animals were in a stage of regression, whereas the mammae of the young virgin rats showed the highest degree of lobulo-alveolar development. The levels of estrogen receptor (ER), epidermal growth factor (EGF) receptor (EGF-R) and GH receptor (GHR) in the mammary glands of the animals were also measured. We found a reduction in the receptor levels for both estrogen and EGF in mammary tissues from parous animals. Receptors for GH were present in normal mammary tissues from both virgin and parous rats. We hypothesize that the reduction in the circulating concentration of GH caused the reduced susceptibility of parous rats to mammary carcinogenesis possibly by decreasing the levels of ER and/or EGF-R in the mammary gland.

Animals↗
PubMed1995

Uptake and distribution of the boron-containing ether lipid B-Et-11-OMe in tumor-bearing mice.

Ether lipids in general are accumulated in tumor tissue with a favorable tumor/healthy tissue ratio. The uptake of the boron-containing analog rac-1-(9-o-carboranyl)nonyl-2-methyl-glycero-3-phosphocholine (B-Et-11-OMe) was studied in C3H mice bearing the murine mammary carcinoma AT17 and in BALB/c mice bearing an osteosarcoma. Boron concentrations of tumor, blood, liver and kidney were followed up to 48 h by inductively coupled plasma emission spectrometry and inductively coupled plasma mass spectrometry. Boron concentration in AT17 mamma carcinoma rose up to 2 mg/kg and the tumor/blood ratio rose to 0.5. The bulk was taken up by the liver. Osteosarcoma did not take up B-Et-11-OMe. This result constitutes a significant contrast to the behavior of published (non-boron-containing) analogs. It is interpreted in terms of critical micellar concentration (CMC). Whereas earlier work with ether lipids was done well below CMC, this study was undertaken above. Further studies will concentrate on syntheses of high CMC analogs.

Adenocarcinoma↗
PubMed1982

129I in Missouri thyroids.

Concentrations of 129I and values of the 129I/127I ration are reported in one sample of indigenous vegetation and in over forty additional individual thyroids of man, wild deer and beef cattle in Missouri. The results of this and other studies in our laboratory indicate the following order for successively lower values of 129I/127I ratios in the local environment: Rain, wild deer, commercial milk, beef cattle and human. The value of the 129I/127I ratio in the single vegetation sample is intermediate to the mean values in wild deer and commercial milk, but well within the range of values observed in both. These results are consistent with a geochemical cycle in which iodine that is enriched in 129I is transported via air into the central U.S. and then diluted with other iodine--especially mineral iodine that is added to the diets of domesticated animals--as the iodine deposited from air moves through the local ecosystem. Differences in the diets of beef and dairy cattle or differences in the biological life-times of iodine in thyroids and mammae, and hence the degree of equilibration with body iodine, may explain the lower value of the 129I/127I ratio in beef thyroids than in milk.

Animals↗
PubMed1996

Soluble CD44 isoforms in serum as potential markers of metastatic gastric carcinoma.

A splice variant of CD44 (exon V4-V7) confers metastatic behavior in a rat carcinoma model; aberrant expression of splice variants has been detected on a variety of human tumor cell lines as well as primary and metastatic human tumors, including lymphomas, carcinomas (colon, thyroid, mamma, bladder), and glioma. We used enzyme-linked immunosorbent assay to determine the concentration of soluble CD44 in the serum samples of 10 normal individuals and 41 patients with various stages of gastric cancer. Soluble CD44S and its isoforms, V5 and V6, were present in the serum of normal individuals (288.53 +/- 18.33, 25.49 +/- 1.70, and 148.32 +/- 3.15 ng/ml, respectively). The concentrations of soluble CD44 V5 and V6 were elevated in patients with advanced gastric carcinoma (69.39 +/- 6.06 and 216.62 +/- 32.98 ng/ml, respectively). Serum CD44 V5 concentrations correlated with the extent of tumor invasion (T), the status of lymph node involvement (N), and distant metastasis (M) (TNM staging) (p < 0.05), whereas CD44S did not. These results suggest that detection of abnormal regulation of CD44 splicing could be helpful in gastric cancer diagnosis and disease evaluation.

Biomarkers, Tumor↗
PubMed2000

The differential time courses of the vasodilator effects of various 1,4-dihydropyridines in isolated human small arteries are correlated to their lipophilicity.

OBJECTIVES: To investigate a possible relationship between the time courses of action of various calcium antagonists and their lipophilicity, characterized as log P-values. METHODS: The functional experiments were performed in vitro in human small subcutaneous arteries (internal diameter 591 +/- 51 microm, n = 7 for each concentration), obtained from cosmetic surgery (mamma reduction and abdominoplasty). The vessels were investigated in an isometric wire myograph. The vasodilator effect of the calcium antagonists was quantified by means of log IC50-values, and the onset of the vasodilator effect for each concentration studied was expressed as time to Eeq90-values (time to reach 90% of the maximal effect). RESULTS: Log IC50-values were -8.46 +/- 0.09, -8.33 +/- 0.25 and -8.72 +/- 0.16 for nifedipine, felodipine and (S)-lercanidipine, respectively (not significant). On average, nifedipine reached time to Eeq90 in 11 +/- 1 min. For felodipine and (S)-lercanidipine the corresponding values were 60 +/- 11 min and 99 +/- 9 min, respectively. The differences between these values were statistically significant (P< 0.01). In spite of these differences in the in-vitro human vascular model, the three calcium antagonists are equipotent with regard to their vasodilator effects. Linear regression analysis of the correlation between the logarithm of the membrane partition coefficient (log P-values) of the calcium antagonists tested [2.50, 4.46 and 6.88 for nifedipine, felodipine and (S)-lercanidipine, respectively] and their respective values found for time to Eeq90 was highly significant. CONCLUSIONS: It appears that a higher log P-value is correlated with a slower onset of action.

Adipose Tissue↗
PubMed2004

Matrix metalloproteinases, tissue inhibitors of metalloproteinases, aminoterminal propeptide of procollagen type III, and hyaluronan in sera and tissue of patients with capsular contracture after augmentation with Trilucent breast implants.

In various fibrotic diseases, matrix metalloproteinases (MMPs) and their natural inhibitors, the tissue inhibitors of metalloproteinases (TIMPs), play an important role. In our study, serum concentrations of MMP-1, MMP-2, MMP-9, TIMP-1, and TIMP-2 were determined by enzyme-linked immunosorbent assay in 17 female patients with Baker grade II (n =9), III (n =7), and IV (n =1) capsular contracture after bilateral cosmetic mamma augmentation with Trilucent implants (AEI, Inc., Caversham, United Kingdom). Samples of capsular tissue for standard histology and immunohistochemistry were obtained from all patients. Sera from 20 female patients who had plastic surgery for reduction mammaplasty were used as the control group. The aminoterminal propeptide of procollagen type III (PIIINP) and hyaluronan were analyzed as markers for fibrogenesis in both groups, too. Statistical analysis was performed using the Mann-Whitney test and Spearman rank correlation. Patients with capsular contracture presented significantly higher concentrations of TIMP-1 and TIMP-2 in their sera than did the control group (p < 0.05), which correlated with Baker grade (r = 0.7 versus r = 0.65; p < 0.05). The concentration of MMP-2 was significantly higher in the sera of patients with capsule fibrosis, whereas there were no significant differences in MMP-1, MMP-9, and PIIINP serum concentrations. Patients with capsule fibrosis had a significantly lower MMP-to-TIMP ratio (1.1 +/- 0.4, p <0.05) than the control group (1.5 +/- 0.4), which correlated with the Baker classification (r =0.7; p <0.05). The hyaluronan serum concentration of patients with capsular contracture was significantly higher (p < 0.05) and correlated with the Baker grade (r = 0.73; p < 0.05), whereas PIIINP showed no difference. In the histologic evaluation, there was a chronic inflammatory reaction in the capsules around the breast implants and refracting material within the substance. Immunohistochemically, TIMP-1 and TIMP-2 showed an intensive accumulation, and MMP-2 showed a local reaction. PIIINP could be detected, too, whereas there was no staining for MMP-1 and MMP-9. The elevated systemic MMP-2 concentration and the local positive staining in the tissue might be due to the chronic inflammatory reaction. Nevertheless, the balance between MMPs and their natural inhibitors is disturbed in patients with capsule contracture. The elevated systemic concentration of TIMPs might be a pathway in the pathogenesis of severe fibrosis after breast augmentation with alloplastic material. Hyaluronan might be a useful marker for early prediction of capsule fibrosis, whereas PIIINP is not useful as a predictor.

Adult↗
PubMed1986

Butcher's wart virus (HPV 7) infections in non-butchers.

The analysis of a total of 654 benign and malignant lesions of the skin, genitalia, lungs and bronchi, intestine, kidneys, bladder, mammae, and of the head and neck region, resulted in the identification of human papilloma virus 7 (HPV 7) infections in 3 individuals. One of these was an ordinary "butcher's wart," whereas the other 2 patients have never been involved in meat-handling or farming. One of the latter revealed extensive verrucosis of hands, feet, axilla, neck, and face, persisting for about 27 years, with new lesions arising in the neck region. Particularly the new lesions showed filiform morphology. The second patient has, for a period of more than 2 years, been showing filiform papillomas in the face which recurred after surgical removal. These 2 patients appear to represent the first cases of HPV 7 infections in non-butchers or non-meathandlers.

Abattoirs↗
PubMed1998

Borrelial lymphocytoma--a historical case.

We here describe a patient with a tick bite in the areola mammae in 1953 followed by erythema migrans. Twenty years later, after another tick bite in the axillary skin, also followed by erythema migrans, a large lymphatic infiltrate developed in the mammary skin, when the margin of the erythema reached the areola. The infiltrate resolved within a year without any therapy. Borrelial DNA was detected by polymerase chain reaction in the paraffin blocks of the lymphatic skin infiltrate. The patient died 9 years later of generalized lymphoma. A similar monoclonal immunoglobulin heavy chain gene rearrangement was detected both in the mammary skin lesion and in the lymphoma specimen.

Animals↗
PubMed1977

A critical review of current methods for induction of parturition in the mare.

The efficacy and safety of oxytocin, dexamethasone and prostaglandin, used alone or in combination as inducing agents, are discussed. It is contended that insufficient evidence exists to support the routine application of any of these methods in practice. Oxytocin has been the most widely used and it is claimed by some to be free from side effects. However, the synthetic prostaglandin analogue, fluprostenol, seems to pose the least risk to the foetus and dexamethasone appears to be either ineffective, or too dangerous to use at all. The main indications for induced foaling are managerial convenience or for research and teaching purposes. There are few clinical indications, although ventral rupture and cases of prolonged gestation have been mentioned by various workers. It is considered that foetal maturity is the pre-requisite before a decision to induce should be made in practice, and 3 criteria are essential: 1) a gestational length of greater than 320 days, 2) substantial mammary development, 3) the presence of colostrum in the mammae.

Animals↗
PubMed1977

Non-neoplastic memmary hypertrophy in the cat associated either with pregnancy or with oral progestagen therapy.

Nine cases of mammary glandular hypertrophy in the cat are described. Three cases occurred in pregnant queens aged between six and eight months and one in an aged non-pregnant queen. The other cases were in neutered animals which had been treated with megestrol acetate for periods of between 14 months and five years. The pregnant queens were spayed and their mammae returned to normal; in the other cats the affected mammary glands were removed surgically. No recurrence occurred in seven cats in which adequate follow-up information was available. The importance of differentiating this benign lesion from mammary neoplasia is discussed.

Animals↗
PubMed1988

[Detection of liver metastases in gynecologic neoplasms by sonography, scintigraphy, computerized tomography and liver enzymes].

In a retrospective study the diagnostic validity of sonography (US), computer-tomography (CT), scintigraphy (SC) and serum alkaline phosphatase (AP) in the detection of liver metastases was evaluated in 929 patients with malignant tumors: ovary (n = 367), mamma (n = 189), endometrium (n = 181), cervix (n = 162), fallopian tube (n = 10), vulva (n = 20). Definitive diagnosis was confirmed by autopsy (n = 51), surgical intervention (n = 297) or follow-up (n = 581). Specificity, sensitivity and overall accuracy of the different methods in the indication of liver metastasis were as follows: CT (n = 58) 81%, 98%, 93%, US (n = 929) 70%, 94%, 90%, SC (n = 512) 66%, 85%, 81%, AP (n = 325) 68%, 93%, 87%. In the last examination period, US and CT reached comparable results. In view of efficiency, AP and US would appear suitable for routine control in gynecological malignancies.

Alanine Transaminase↗
PubMed1989

[Additional treatment with Esberitox N in patients with chemo-radiotherapy treatment of advanced breast cancer].

A study was made at random on 70 female patients with advanced mamma carcinoma to find out whether an additional treatment with Esberitox N prevents or diminishes the sideeffects of a combined chemo-radiation therapy. As it could not be excluded that the efficacy of Esberitox N on previous affliction by haematotoxic therapy would probably differ, the study was made on various groups of risk. The parameter to form an opinion were either the peripheral hematosis or the appearance of infections. It turned out, however, that Esberitox N proved only effective if there was only a minor damage of the bonemarrow. The efficacy seems to promote the recuperation of the haemapoetic system. There was no significant frequency of infections by the additional Esberitox N treatment, yet in tendency favourably influenced.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1977

Use of partial mammectomy to produce malnutrition in the rat.

Partial mammectomy in the Sprague-Dawley rat was studied as a method for producing malnutrition without accompanying differences in litter size. Throughout the litter period, experimental litters weighed less than controls. No difference in the rate of development of various anatomical landmarks was noted with this degree of malnutrition. Maternal behavior in the nursing dam was measured with respect to nursing, nesting, and retrieval behavior. Mammectomized subjects built and maintained better nests and spent more time in a nursing posture, but were less likely to retrieve their litter than were control animals. At weaning the intact mammae of the experimental animals were larger than those of control animals.

Animals↗
PubMed1988

Comparison of histology and clinical variables to DNA ploidy in canine mammary tumors.

Flow cytometric DNA analysis was done on 132 canine mammary tumors from 99 dogs to evaluate the relation to histology and to clinical staging. Seventy-one tumors (54%) were histologically malignant; 38 (54%) of these were aneuploid and 33 (46%) were diploid. Fifty-two (39%) tumors were histologically benign, of which 45 (87%) were diploid and seven (13%) aneuploid. There were nine dysplastic mammae (7%); two were aneuploid and the rest diploid. DNA indices varied from 0.72 to 2.35. Of 58 mammary carcinomas, 25 (43%) were diploid and 33 (57%) were aneuploid (of the latter, 16 showed hypodiploidy and 17 hyperdiploidy with a predominance between DNA index 1.10 and 1.50). Three tumors (two carcinomas and one malignant mixed tumor) were multiploid with two aneuploid cell populations. The histological type varied within eight tumors, and in four of these the DNA index also varied. DNA indices varied within three tumors with uniform morphology. No correlation was found between DNA index and age of the dogs, nor between DNA index and tumor size. No significant differences were found between DNA index and histology, tumor growth pattern, or tumor location. Benign tumors were smaller than carcinomas, which were smaller than malignant mesenchymal tumors. Tumors growing adherent to the skin were larger than those not adherent to the skin. The regional lymph nodes were examined in 33 cases. No significant difference between the mean DNA index and presence of lymph node metastasis was found. These results show the possibility of using flow cytometry for DNA analysis in canine mammary tumors.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1996

Experimental infection of pregnant cattle with the vaccine candidate Brucella abortus strain RB51: pathologic, bacteriologic, and serologic findings.

To determine the placental tropism and abortigenicity of the vaccine candidate Brucella abortus strain RB51 (SRB51), a rough mutant of the virulent strain 2308, ten Polled Hereford heifers were inoculated intravenously in the 6th month of gestation. Heifers were euthanatized and examined at postinoculation week (PIW) 8 (n = 5) or at full term (n = 5). Four of five infected heifers sampled at PIW 8 and three of four infected heifers at term had placentitis, whereas reproductive tissues of three normal cows used for comparison had no placentitis. Numerous macrophages, immunoreactive for SRB51 antigen, as well as neutrophils, fibrin, and cell debris filled the arcade zone between chorion and maternal septae. Trophoblastic epithelium of the placentomal arcade zone had intracellular bacteria that were immunoreactive for SRB51 antigen. The tips of maternal septa had a lymphoplasmacytic infiltrate with small multifocal erosions and ulcerations of maternal epithelium. SRB51 was cultured from all tissues in which lesions were seen. Placentae of one cow from each group had no placentitis and contained no SRB51. In mammae, interstitial lymphoplasmacytic infiltrates and suppurative infiltrates within alveoli and intralobular ductules were seen in two of five heifers at PIW 8. SRB51 was cultured from liver, spleen, lung, and bronchial lymph nodes in four of five calves at PIW 8 and three of four full-term calves, but no lesions were seen. One near-term heifer had disseminated infection, placentitis, and lymphoplasmacytic endometritis, and delivered a premature weak calf. These results establish that SRB51 is less abortifacient than previously published reports with strain 19, in that only one of four heifers delivered prematurely following intravenous inoculation with SRB51, whereas intravenous inoculation with strain 19 leads to 100% abortion. However, it also shows that SRB51 can infect the bovine placenta, mammary gland, and fetus, can induce placentitis, and, in some cases, can lead to preterm expulsion of the fetus.

Abortion, Veterinary↗