RNA-based determination of ESR1 and HER2 expression and response to neoadjuvant chemotherapy.

BACKGROUND: Hormone and human epidermal growth factor receptor 2 (HER2) receptors are the most important breast cancer biomarkers, and additional objective and quantitative test methods such as messenger RNA (mRNA)-based quantitative analysis are urgently needed. In this study, we investigated the clinical validity of RT-PCR-based evaluation of estrogen receptor (ESR1) and HER2 mRNA expression. PATIENTS AND METHODS: A total of 1050 core biopsies from two retrospective (GeparTrio, GeparQuattro) and one prospective (PREDICT) neoadjuvant studies were evaluated by quantitative RT-PCR for ESR1 and HER2. RESULTS: ESR1 mRNA was significantly predictive for reduced response to neoadjuvant chemotherapy in univariate and multivariate analysis in all three cohorts. The complete pathologically documented response (pathological complete response, pCR) rate for ESR1+/HER2- tumors was 7.3%, 8.0% and 8.6%; for ESR1-/HER2- tumors it was 34.4%, 33.7% and 37.3% in GeparTrio, GeparQuattro and PREDICT, respectively (P < 0.001 in each cohort). In the Kaplan-Meier analysis in GeparTrio patients with ESR1+/HER2- tumors had the best prognosis, compared with ESR1-/HER2- and ESR1-/HER2+ tumors [disease-free survival (DFS): P < 0.0005, overall survival (OS): P < 0.0005]. CONCLUSIONS: Our results suggest that mRNA levels of ESR1 and HER2 predict response to neoadjuvant chemotherapy and are significantly associated with long-term outcome. As an additional option to standard immunohistochemistry and gene-array-based analysis, quantitative RT-PCR analysis might be useful for determination of the receptor status in breast cancer.

Radioautographic visualization of differences in the pattern of [3H]uridine and [3H]orotic acid incorporation into the RNA of migrating columnar cells in the rat small intestine.

The epithelium of rat small intestine was radioautographed to examine whether RNA is synthesized by the salvage pathway as shown after [3H]uridine injection or by the de novo pathway as shown after [3H]orotic acid injection. The two modes of RNA synthesis were thus investigated during the migration of columnar cells from crypt base to villus top, and the rate of synthesis was assessed by counting silver grains over the nucleolus and nucleoplasm at six levels along the duodenal epithelium--that is, in the base, mid, and top regions of the crypts and in the base, mid, and top regions of the villi. Concomitant biochemical analyses established that, after injection of either [5-3H]uridine or [5-3H]orotic acid: (a) buffered glutaraldehyde fixative was as effective as perchloric acid or trichloracetic acid in insolubilizing the nucleic acids of rat small intestine; (b) a major fraction of the nucleic acid label was in RNA, that is, 91% after [3H]uridine and 72% after [3H]orotic acid, with the rest in DNA; and (c) a substantial fraction of the RNA label was in poly A+ RNA (presumed to be messenger RNA). In radioautographs of duodenum prepared after [3H] uridine injection, the count of silver grains was high over nucleolus and nucleoplasm in crypt base cells and gradually decreased at the upper levels up to the villus base. In the rest of the villus, the grain count over the nucleolus was negligible, while over the nucleoplasm it was low but significant. After [3H]-orotic acid injection, the number of silver grains over the nucleolus was negligible at all levels, whereas over the nucleoplasm the number was low in crypt cells, but high in villus cells with a peak in mid villus. The interpretation is that, except for a small amount of label incorporated into DNA from either precursor by crypt cells, the bulk of the label is incorporated into RNA as follows. In the crypts, cells make almost exclusive use of uridine, that is, of the salvage pathway, for the synthesis of ribosomal RNA in the nucleolus and of messenger and transfer RNA in the nucleoplasm. However, when cells pass from crypt to villus, they mainly utilize orotic acid--i.e., the de novo pathway--for the synthesis of messenger and transfer RNA within the nucleoplasm.

[HES 130 as a continuous supplement for organ culture of human corneas?]. / HES 130 als Dauerzusatz in der Organkultur humaner Hornhäute?

BACKGROUND: During organ culture, corneas swell and have to be de-swollen prior to transplantation using a medium. Both swelling and de-swelling lead to increased endothelial cell loss. Furthermore, dextran, the standard de-swelling substance, is toxic and deposited in the cornea. Whether HES is suitable as a continuous supplement for organ culture and keeps the corneas thin was investigated in this study. METHODS: Forty-five human corneas were stored in MEM plus 7.5% HES 130 for 7, 15, 21, and 28 days. Endothelial cell count was determined before and after organ culture. After storage the water content and amount of adenosine phosphates were measured. RESULTS: Water content was 82.29% after 7 days, 82.98% after 15 days, 81.15% after 21 days, and 83.21% after 28 days. The endothelial cell count decreased by 1.20% after 7 days, 0.60% after 15 days, 4.29% after 21 days, and 6.89% after 28 days. ATP was 0.159 micromol/g dry weight after 28 days of organ culture. CONCLUSION: The water content of corneas stored in HES-containing medium remained constant even after 21 days of organ culture. Endothelial cell loss was less than in comparable studies using a standard medium containing 10% FCS; however, ATP concentrations were worse. HES 130 keeps the corneas thin and can be used as a continuous supplement. It facilitates the handling of corneas during organ culture, making dehydration before transplantation unnecessary, and thus also reduces potential stress factors for the cornea.

Effects of 1,2-dimethylhydrazine on the number of epithelial cells present in the villi, crypts, and mitotic pool along the rat small intestine.

Young adult male rats received 1,2-dimethylhydrazine (25 mg/kg) twice weekly for 2 months and once a week thereafter for up to 6 months. Histological samples of duodenum, jejunum, and upper, mid-, and terminal ileum were prepared from groups killed at each month. Using cell counts, the average number of epithelial cells was determined per representative section of villi and crypts and was used as an index of villus size and crypt size, respectively. The average number of mitotic figures in representative crypt sections was also determined. All three parameters increased during treatment, and the increments showed a specific pattern in relation to time and intestinal region. Mitotic number showed a consistent change all along the small intestine: close to 20% rise by 3 months; decrease to near control level by the fourth month; and a rise thereafter. Probably, a systemic stimulation of mitotic activity by 1,2-dimethylhydrazine took place. The crypt size index changed similarly, showing highly significant correlation with mitotic number. This correlation indicated that average mitotic time and cell cycle time remained unchanged and the number of divisions increased in progenitor cells. Calculations showed that only a fraction of the progenitor cells was involved. These were probably "initiated" cells. There was a net increase of initiated cell numbers with time, but a sharp drop at 4 months indicated that there is a mechanism inducing a regression of the initiated cell population. Villus size increased linearly in the duodenum and jejunum. In the ileum, there was also a net increase but with some initial fluctuation. In general, villus size seemed to increase so as to maintain a fairly stable turnover time. This would mean that the increased mitotic activity was balanced by increasing differentiation.

Morphometric study of the rat duodenal epithelium during the initial six months of 1,2-dimethylhydrazine carcinogenesis.

Male Wistar rats weighing 100 g received 1,2-dimethylhydrazine (25 mg/kg) s.c.) twice a week for 2 months and once a week thereafter for an additional 4 months. Groups of four to six rats were sacrificed monthly. Paraffin sections of duodenum were prepared and stained with periodic acid-Schiff and hematoxylin for cell counts and with toluidine blue for measuring nucleolar area. As an index of villus size and crypt size, the mean number of epithelial cells per representative sections of villi and crypts were used. Mitotic activity was assessed by counting the mean number of mitotic figures per representative crypt section. Nucleolar area was assessed from the analysis of drawn (camera lucida) images of nucleoli of columnar cells at six levels of the epithelium: lower, mid, and upper parts of the crypts and villi. Villus size increased progressively during the 6-month treatment, from 272 +/- 2 (S.E.) to 349 +/- 8. Crypt size increased from 118 +/- 2 in a wavy fashion, showing maximum (139 +/- 5, 143 +/- 2) at 3 and 6 months and minimum (123 +/- 3, 127 +/- 1) at 1 and 4 months. Mitotic number displayed a similar pattern of increase so that the percentage of mitotic figures in the crypts (mitotic index) remained constant (about 5%) in control and experimental animals. Nucleolar area in the controls decreased with age from 4.2 +/- 0.08 sq micron in lower crypt at 1 month to 2.8 +/- 0.04 sq micron at 6 months. During 1,2-dimethylhydrazine treatment, lower crypt nucleoli increased to 4.5 +/- 0.12 sq micron after 3 months and decreased slightly thereafter, reaching 4.0 +/- 0.14 sq micron by the sixth month. The nucleoli furthermore decreased gradually along epithelium (nucleolar compaction) by an average of 0.23% per cell position in control as well as treated animals. It appeared, then, that the main effect of 1,2-dimethylhydrazine was the enlarging of the four parameters measured. This effect seemed to relate in some manner to tumor formation as all the enlargements were attenuated in intestinal tissue adjacent to tumors.

Ambiguity in sentence processing.

As listeners and readers, we rarely notice the ambiguities that pervade our everyday language. When we hear the proverb `Time flies like an arrow' we might ponder its meaning, but not the fact that there are almost 100 grammatically permissible interpretations of this short sentence. On occasion, however, we do notice sentential ambiguity: headlines, such as `Two Sisters Reunited After 18 Years in Checkout Counter', are amusing because they so consistently lead to the unintended interpretation (presumably, the sisters did not spend 18 years at the checkout). It is this consistent preference for one interpretation-and one grammatical structure-rather than another that has fuelled research into sentence processing for more than 20 years. Until relatively recently, the dominant belief had been that these preferences arise from general principles that underlie our use of grammar, with certain grammatical constructions being preferred over others. There has now accrued, however, a considerable body of evidence demonstrating that these preferences are not absolute, but can change in particular circumstances. With this evidence have come new theories of sentence processing, some of which, at first glance, radically question the standard notions of linguistic representation, grammar and understanding.

Incremental interpretation at verbs: restricting the domain of subsequent reference.

Participants' eye movements were recorded as they inspected a semi-realistic visual scene showing a boy, a cake, and various distractor objects. Whilst viewing this scene, they heard sentences such as 'the boy will move the cake' or 'the boy will eat the cake'. The cake was the only edible object portrayed in the scene. In each of two experiments, the onset of saccadic eye movements to the target object (the cake) was significantly later in the move condition than in the eat condition; saccades to the target were launched after the onset of the spoken word cake in the move condition, but before its onset in the eat condition. The results suggest that information at the verb can be used to restrict the domain within the context to which subsequent reference will be made by the (as yet unencountered) post-verbal grammatical object. The data support a hypothesis in which sentence processing is driven by the predictive relationships between verbs, their syntactic arguments, and the real-world contexts in which they occur.

Two modes of transfer in artificial grammar learning.

Participants can transfer grammatical knowledge acquired implicitly in 1 vocabulary to new sequences instantiated in both the same and a novel vocabulary. Two principal theories have been advanced to account for these effects. One suggests that sequential dependencies form the basis for cross-domain transfer (e.g., Z. Dienes, G. T. M. Altmann, & S. J. Gao, 1999). Another argues that a form of episodic memory known as abstract analogy is sufficient (e.g., L. R. Brooks & J. R. Vokey, 1991). Three experiments reveal the contributions of the 2. In Experiment 1 sequential dependencies form the only basis for transfer. Experiment 2 demonstrates that this process is impaired by a change in the distributional properties of the language. Experiment 3 demonstrates that abstract analogy of repetition structure is relatively immune to such a change. These findings inform theories of artificial grammar learning and the transfer of grammatical knowledge.

Evaluation of countercurrent immunoelectrophoresis as a diagnostic tool in bacterial otitis media.

The diagnostic efficiency of countercurrent immunoelectrophoresis (CIE) and correlation of clinical and bacteriological data were studied in 184 middle ear effusions (MEE) from 125 patients diagnosed as acute primary, acute recurrent otitis media, and chronic MEE. Significant differences between stage of otitis media (OM), patient age and type of MEE were found. Streptococcus pneumoniae was isolated in 22% of the overall MEE, 26% in the acute primary and 26% in acute recurrent OM. Among the S. pneumoniae isolates, 82.5% were detected by CIE and culture, 15% only by culture and 2.5% only by CIE. Most S. pneumoniae isolates identified by culture and CIE were in a purulent MEE (64%) while those identified by culture only were in a mucoid MEE (67%). Haemophilus influenzae overall incidence was 29%; 13% of them produced beta-lactamase and only 4% were serotype b. The H. influenzae serotype b strains were detected by culture as well as CIE, so the overall effectiveness of CIE for S. pneumoniae and H. influenzae serotype b was 19.5% and 22 to 23% for acute primary and recurrent OM. Sterile cultures were obtained in 37.5% of the overall MEE. The CIE seems to be a useful screening test, culture remaining the method for definitive bacterial identification in MEE.

[Results of radical Wertheim's operation 1964 to 1972 (author's transl)]. / Ergebnisse der Wertheimschen Radikaloperation in den Jahren 1964 bis 1972.

In the years 1964 to 1972 239 women with cervical carcinoma were treated by extended abdominal radical (Wertheim's) operation at the II. Department of Obstetrics and Gynaecology, University of Vienna. The total number of invasive cervical carcinomas is decreasing in comparison with previous years, but the figure of 40% cases treated surgically remains stable. 69.5% of the women are alive after a 5-year observation period, the figure for stage Ib being 71.4% and for stage II 55.2%. In cases with positive lymph nodes an improved 5-year survival rate is demonstrated and the total number of patients who died of carcinoma is reduced in comparison with the respective figures for previous years.

[Ultrasonic diagnosis of ovarian tumours (author's transl)]. / Ultraschalldiagnostik von Adnextumoren.

Precise knowledge of the echoanatomy of the normal female pelvis is a prerequisite in the diagnosis of ovarian tumours. Diagnostic criteria for the differentiation between cystic and solid lesions are discussed. The echographic appearance of uni-, multilocular, dermoid and endometriosis cysts and of ovarian cancer is described. 183 ovarian tumours were examined in the period from January 1st, 1976 until December 31st, 1977 and the preoperative diagnosis compared with the findings at operation, both macroscopic and histological. 158 cases (86.3%) were correctly diagnosed. In 13 cases (7.1%) the diagnosis was only partially correct. An incorrect ultrasonic diagnosis was made in only 12 cases (6.6%). Out of 183 cases the side of the lesion was correctly diagnosed in 128 patients (70.0%). Localization of the tumour could not be achieved or was totally wrong in 19 cases (10.4%). Estimated tumour dimensions concurred with the macroscopic measurements of the specimen in 90.7%. A false-positive result was recorded in 7 cases (3.8%), mostly due to a full bowel simulating a cyst of barely 4 cm in diameter. In 5 cases (2.7%) ultrasonic examination failed to detect the existence of a tumour. 2 of these tumours were follicular cysts under 4 cm in diameter. 2 ovarian cancers were missed. In 1 case the ovary was only slightly enlarged and the diagnosis was made only by frozen-section histology. In the second case the tumour was erroneously interpreted as originating from the sigmoid colon. Ultrasonic diagnosis is recommended for the detection and differentiation of ovarian tumours on the basis of the good results achieved.

[The clinical efficacy of the copper-containing intrauterine devices Nova T and Copper-T-200 Ag in a randomized study (author's transl)]. / Die klinische Wirksamkeit der kupferhältigen Intrauterinpessare Nova T und Kupfer-T-200 Ag im randomisierten Vergleich.

In a randomized study the clinical efficacy of the traditional intrauterine device. Copper-T-200 Ag was compared with the recently developed model Nova T. Altogether 93 women with regular menstrual cycles were examined; 45 women had a Nova T inserted and 48 the traditional Copper-T-200 Ag. The biosocial data of the women in the two groups did not differ from one another. No pregnancies occurred in either group within the first 12 months. Expulsion occurred in 2 women with Nova T and removal of the device had to be undertaken for bleeding and pain in 3 women with a Copper-T-200 Ag. At 12 months the two models did not differ in the number of recorded incidents. The reliability of randomized studies is discussed, as well as problems arising from the fact that the mode of insertion is not identical with both devices and requires practice in the case of the new model.

PIP: In a randomized study, the clinical efficacy of the Copper T-200 Ag IUD was compared to the recently developed model Nova T. 93 women with regular menstrual cycles were examined; 45 women had a Nova T inserted and 48 received the traditional Copper T-200 Ag. The biosocial data of the women in the 2 groups did not differ. No pregnancies occurred in either group within the first 12 months. Expulsion occurred in 2 women with the Nova T and removal of the device was undertaken for bleeding and pain in 3 women with the Copper T-200 Ag. At 12 months, the 2 models did not differ in the number of recorded incidents. The reliability of randomized studies is discussed, as well as problems arising from the fact that the mode of insertion is not identical with both devices and requires practice in the case of the newer model. (author's)