Principal component analysis of three-dimensional face shape: Identifying shape features that change with age.

BACKGROUND: The types of shape feature that constitutes a face have not been comprehensively established, and most previous studies of age-related changes in facial shape have focused on individual characteristics, such as wrinkle, sagging skin, etc. In this study, we quantitatively measured differences in face shape between individuals and investigated how shape features changed with age. METHODS: We analyzed three-dimensionally the faces of 280 Japanese women aged 20-69 years and used principal component analysis to establish the shape features that characterized individual differences. We also evaluated the relationships between each feature and age, clarifying the shape features characteristic of different age groups. RESULTS: Changes in facial shape in middle age were a decreased volume of the upper face and increased volume of the whole cheeks and around the chin. Changes in older people were an increased volume of the lower cheeks and around the chin, sagging skin, and jaw distortion. CONCLUSION: Principal component analysis was effective for identifying facial shape features that represent individual and age-related differences. This method allowed straightforward measurements, such as the increase or decrease in cheeks caused by soft tissue changes or skeletal-based changes to the forehead or jaw, simply by acquiring three-dimensional facial images.

Abnormal FHIT transcripts in human breast carcinomas: a clinicopathological and epidemiological analysis of 61 Japanese cases.

Deletions in the short arm of chromosome 3 have been found in various human cancers, including breast cancer. Recently, the FHIT (fragile histidine triad) gene was identified at 3p14.2 as a candidate tumor suppressor gene. We examined the abnormal transcripts of the FHIT gene in 61 Japanese primary breast cancer specimens and found that 23 (38%) of them exhibited abnormalities, about half of which were categorized into two types of aberrant transcripts. Sequence analysis of these aberrant transcripts revealed the absence of exons 5-7 (type I) and exons 5-8 (type II). Clinicopathological and epidemiological analysis of patients showed that the abnormal FHIT transcripts were not associated with age, tumor-node-metastasis classification, tumor size, estrogen receptor and progesterone receptor status, local metastasis, family history of breast cancer, or lifestyle factors of patients, including cigarette smoking and alcohol consumption. On the other hand, we found that the abnormal transcripts of type I and type II were associated with the incidence of bilateral breast cancer and that decreased frequency of childbirth was also associated with FHIT abnormalities.

Spontaneous long-term hyperglycemic rat with diabetic complications. Otsuka Long-Evans Tokushima Fatty (OLETF) strain.

A spontaneously diabetic rat with polyuria, polydipsia, and mild obesity was discovered in 1984 in an outbred colony of Long-Evans rats, which had been purchased from Charles River Canada (St. Constant, Quebec, Canada) in 1982. A strain of rats developed from this rat by selective breeding has since been maintained at the Tokushima Research Institute (Otsuka Pharmaceutical, Tokushima, Japan) and named OLETF. The characteristic features of OLETF rats are 1) late onset of hyperglycemia (after 18 wk of age); 2) a chronic course of disease; 3) mild obesity; 4) inheritance by males; 5) hyperplastic foci of pancreatic islets; and 6) renal complication (nodular lesions). Histologically, the changes of pancreatic islets can be classified into three stages: 1) an early stage (6-20 wk of age) of cellular infiltration and degeneration; 2) a hyperplastic stage (20-40 wk of age); and 3) a final stage (at > 40 wk of age). These clinical and pathological features of disease in OLETF rats resemble those of human NIDDM.

New inbred strain of Long-Evans Tokushima lean rats with IDDM without lymphopenia.

Spontaneously diabetic rats with remarkable polyuria, polyphagia, and polydipsia were discovered in 1983 in an outbred colony of Long-Evans rats purchased from Charles River Canada in 1982. They have since been maintained at the Tokushima Research Institute (Otsuka Pharmaceutical, Tokushima, Japan). A strain of rats (Long-Evans Tokushima Lean [LETL]) with diabetes was bred from these rats. The characteristic features of the disease in LETL rats are 1) sudden onset of polyuria, polyphagia, hyperglycemia, and weight loss; 2) no sex differences in the rate of onset or severity; 3) lymphocyte infiltration into islets followed by destruction of beta-cells and disappearance of lymphocytes at the onset of diabetes; 4) no significant T lymphopenia; 5) lymphocyte infiltration into the salivary glands and lacrimal glands; and 6) at least two recessive genes involved in the pathogenesis of insulitis, one of which is closely linked with RT1u. These characteristics closely resemble those of human insulin-dependent diabetes mellitus (IDDM). Results suggest that the LETL rat is a useful animal model for analysis of genetic and immunologic factors relating to the pathogenesis of human IDDM.

Unargued issues on the pathological assessment of response in primary systemic therapy for breast cancer.

PURPOSE: The role of primary systemic therapy (PST) in the treatment of operable breast cancer is currently under intensive investigation in the hope of allowing greater conservation of the breast, and emerging evidence suggests that induction of a pathological complete response (pCR) is at least, to some extent, predictive of long-term clinical response. In this review, we highlight the issues of pathologic evaluation after PST. METHODS: We performed a computer-assisted MEDLINE search, and additional references were found in the bibliographies of these articles. RESULTS: So far, several grading classifications are used to assess pathologic responses after PST, and pCR rates vary from 1% to 54.7% according to the PST regimens employed. However, the term "pCR" has not been applied in a consistent, standardized manner, and the pCR rates appear to depend not only on the differences in the definition of pCR, but also on the extent of tissue sampling and the techniques used for pathologic examination. So far, only limited information is available about the reliability and validity of the definition of pathologic responses. CONCLUSION: Assessment of pCR needs to be standardized, and each grading system should be verified for reliability and validity. As a lack of standard for tumor processing and evaluation may result in considerable fluctuation of pCR rates between trials, we should take into account the differences in the definition of pathologic response when comparing the results of PST.

Postsurgical oral administration of uracil and tegafur inhibits progression of micrometastasis of human breast cancer cells in nude mice.

We recently established a metastasis model in nude mice using the MKL-4 cell line, a contransfectant of the MCF-7 human breast cancer cell line with fgf-4 and lacZ in which micrometastases in several organs can be quantitatively observed. First, to develop a new postsurgical metastasis model, we investigated the timing of occurrence of micrometastasis and the influence of tumor removal on the progression of micrometastasis in this model. Micrometastases into lymph nodes and lungs were detected 3 weeks after the cell injections. Tumor removal 3 weeks after the injections significantly enhanced the progression of micrometastasis into lymph nodes and bone. Second, to study the effect of a mixed compound, UFT (a molar ratio of uracil:tegafur of 4:1), which has been widely used in the postsurgical adjuvant setting in Japan, 15 or 20 mg/kg UFT were administered p.o. for 4 weeks to tumor-bearing mice or to mice in which transplanted tumors were resected 3 weeks after the injections. Either dose of UFT significantly inhibited the tumor growth as well as the progression of micrometastasis into lymph nodes, lungs, liver, and brain. In addition, enhanced progression of micrometastasis in all explored organs by the tumor removal was significantly inhibited by the administration of either dose of UFT. In conclusion, this new postsurgical metastasis model may be useful for evaluating the efficacy of agents used in the postoperative adjuvant setting. UFT may be an effective drug for inhibiting the progression of micrometastasis after surgery.

Invasion-inhibiting factor 2-albumin conjugate inhibits invasion and spontaneous metastasis of MKL-4 human breast cancer cells transplanted into female nude mice.

Invasion-inhibiting factor 2 (IIF-2) and its albumin conjugate have been reported to inhibit spontaneous metastasis of highly metastatic cancer cells with no effect on primary tumor growth. To confirm the inhibitory effects of the IIF-2 conjugate on tumor invasion and spontaneous metastasis, we administered the conjugate intra-peritoneally (i.p.) to female nude mice bearing transplanted tumors with MKL-4 cells, which are MCF-7 human breast cancer cells cotransfected with fibroblast growth factor 4 and lacZ. Neither 10 nor 20 mg/kg doses of the conjugate caused any inhibition of primary tumor growth, but 20 mg/kg significantly inhibited tumor invasion and spontaneous metastasis. Tumor invasion was measured by a novel computer-assisted image analysis. Spontaneous microscopic metastases into lymph nodes and distant organs were measured by whole organ staining for beta-galactosidase activity and observed with a dissecting microscope. The dose of 10 mg/kg significantly inhibited tumor invasion but not metastasis. Interestingly, the number of factor VIII-positive microvessels in the tumors was not reduced by treatment at either dose level. These findings suggest that the anti-invasive effect of the IIF-2 conjugate may reduce both lymphatic and hematogenous metastases in this MKL-4 metastasis model without affecting angiogenesis.

Ovarian hormone receptors in human mammary stromal cells.

Mesenchymal cells of the rodent breast express both estrogen and progesterone receptors. Searches for these molecules in the human breast have yielded conflicting results. Following immunohistochemical staining of samples of normal human breast tissue, the authors detected estrogen receptor alpha protein and progesterone receptor protein in extralobular (non-specialized) fibroblasts and estrogen receptor alpha protein in adipocytes. Tissues from young teenage girls and pregnant women contained the greatest number of receptor positive fibroblasts. These observations confirm prior reports of the presence of ovarian hormone receptors in mammary fibroblasts. The findings also illustrate similarities in the organization of the rodent and human breasts and thereby suggest that regulation of the gland by ovarian hormones involves similar mechanisms in both species.

Combined effects of docetaxel and fluoropyrimidines on tumor growth and expression of interleukin-6 and thymidine phosphorylase in breast cancer xenografts.

PURPOSE: Although several combination treatments with docetaxel and other antitumor agents have been tested in experimental and clinical studies, their synergistic effects are still ill-defined. The degree of synergism between docetaxel and two oral fluoropyrimidines, tegafur and 5'-deoxy-5-fluorouridine (5'-dFUrd), was investigated in the KPL-4 human breast cancer xenograft model. METHODS: Because this KPL-4 cell line secretes interleukin-6 (IL-6) and induces cachexia, the effects of the combined treatment on serum IL-6 levels and cachectic markers were investigated. In addition, the expression levels of thymidine phosphorylase (dThdPase), a key enzyme for converting 5'-dFUrd to 5-fluorouracil, were determined. Female nude mice bearing KPL-4 tumors were treated orally with 5'-dFUrd (60 mg/kg, five times a week) or tegafur (100 mg/ kg, five times a week) and by intraperitoneal injection of docetaxel (5 or 10 mg/kg, once a week). RESULTS: Although docetaxel (5 mg/kg) alone did not decrease either tumor growth or serum IL-6 levels, docetaxel (5 mg/kg) plus 5'-dFUrd or tegafur enhanced tumor growth inhibition and decreased serum IL-6 levels more than 5'-dFUrd or tegafur alone. Docetaxel (5 mg/kg) alone slightly increased the percentage of dThdPase-positive tumor cells, but the combined treatment with docetaxel plus 5'-dFUrd or tegafur significantly decreased the percentage of dThdPase-positive cells in the KPL-4 tumors. CONCLUSION: These findings indicate that docetaxel may stimulate dThdPase expression in tumor tissues and may enhance the antitumor activity of oral fluoropyrimidines. In addition, combined treatment with docetaxel and oral fluoropyrimidines may decrease serum IL-6 levels and may ameliorate IL-6-induced cancer cachexia.

Tumorigenesis in F1 offspring mice following paternal 12.5 cGy 252Cf fission neutron irradiation.

Experiments were conducted to determine whether following genetic damage at germ cell stages induced by paternal exposure to 252Cf fission neutron could lead to tumorigenesis in the offspring. Seven-week-old C3H/HeNCrj male mice were irradiated with 252Cf fission neutrons, at doses of 0 and 12.5 cGy and were mated with nine-week-old C57BL/6NCrj females two weeks after the exposure. Three weeks later, it was found that the proportion of abnormal sperm in the 12.5 cGy-irradiated males was higher than that of 0 cGy-irradiated group. Embryo lethality among the F1 offspring was also found to be higher in the 12.5 cGy group than in the 0 cGy group, while the incidence of liver tumors among the F1 offspring increased in males only. These results suggest that the paternal 12. 5 cGy radiation exposure may have caused genetic transmission of liver tumor-associated traits, which is in line with findings that show steep increase in incidence of tumorigenesis in B6C3F1.

Enhancement of immunohistochemical reactivity for thymidine phosphorylase in breast carcinoma cells after administration of docetaxel as a neoadjuvant chemotherapy in advanced breast cancer patients.

For the purpose of demonstrating possible effects of docetaxel on thymidine phosphorylase (TP) activity in human breast carcinoma, we examined breast carcinoma tissues pre- and post-administration of docetaxel, by an immunohistochemical method using an anti-TP monoclonal antibody. Eight patients with advanced breast carcinoma were initially treated with 3 cycles of 60 mg/m2 of docetaxel once every 3 weeks after incisional biopsy of tumors, and following 3 cycles of docetaxel, they underwent mastectomy with axillary dissection. Grades of immunohistochemical reactivity for TP of carcinoma cells in pre- and post-treatment specimens were compared. Five biopsy specimens (62.5%) were positive for TP. After administration of docetaxel, 6 of 8 cases (75.0%) revealed significant enhancement of reactivity for TP. Increased reactivity was recognized diffusely as well as focally in carcinoma tissues. From these results, we believe that administration of docetaxel to breast cancer patients evokes enhancement of immunohistochemical reactivity for TP in breast carcinoma cells in situ. Furthermore, we consider that docetaxel treatment might improve efficacy of additional doxifluridine and capecitabine therapy.

Relationship between existence of lymphatic invasion in peritumoral breast tissue and presence of axillary lymph node metastasis in invasive ductal carcinoma of the breast.

Specimens obtained from 92 patients with invasive ductal carcinoma of the breast by quadrantectomy and axillary lymph node dissection were examined to evaluate the relationship between existence of lymphatic invasion in peritumoral breast tissue and presence of axillary lymph node metastasis. The number of lymphatic invasions was classified into 4 groups (ly0-3) by counting the number of peritumoral lymphatic invasions. In addition, immunohistochemistry for cytokeratin was performed to locate micrometastasis in the dissected lymph nodes. Thirty-seven (40.2%) of 92 cases had foci of lymphatic invasion and 29 (31.5%) cases revealed lymph node metastasis on initial routine examination. The rate of diagnosis of lymph node metastasis assessed by the existence of lymphatic invasion had an accuracy of 84.8%, a sensitivity of 89.7% and a specificity of 82.5%. On the other hand, all 3 cases (4.8%) with micrometastasis detected by immunohistochemistry for cytokeratin, showed lymphatic invasion. The rate of diagnosis after detection of micrometastasis increased and exhibited 88.0% accuracy. In addition, the rate of prediction of lymph node metastasis in cases with tumor larger than 15 mm was also high, and its accuracy was 88.2%. These results suggest that the assessment of peritumoral lymphatic invasion is very useful for predicting the presence of axillary lymph node metastasis including micrometastasis. They also suggest that excision specimens should be examined for lymphatic invasion, and that the results of the examination might be necessary to pick up false-negative cases and those at high risk for lymph node metastasis among patients who have not undergone lymph node dissection based on the result of sentinel lymph node biopsy.

Effects of azoxymethane on X-ray induced intestinal metaplasia in Donryu rats.

The present study was undertaken to determine the effect of azoxymethane (AOM) administration on intestinal metaplasia induced by X-irradiation in male Donryu rats. Five-week-old animals were X-irradiated with two doses of 10 Gy each at a 3-day interval or three X-ray doses of 10 Gy at a 2-day interval and then received AOM injections i.m. at a dose of 15 mg/kg body weight once weekly for 3 weeks, 6 months after irradiation. Alkaline phosphatase positive foci were decreased after AOM treatments, but aberrant crypt like-foci appeared within areas of intestinal metaplasia. In contrast no induction was observed in normal-appearing gastric mucosa.

The early phase of colon tumorigenesis induced by dimethylhydrazine in ICR mice.

In order to study the influence of fiber supplements on dimethylhydrazine induction of colon tumorigenesis six-week-old CD1 (ICR): Crj mice were injected i.m. at a dimethylhydrazine (DMH) dose 10 mg/kg body weight once weekly for 10 weeks with or without dietary supplementation with 3% polydextrose, lactosucrose or cellulose, or 3% polydextrose and 3% cellulose in combination. There were no significant differences in colon tumor induction among the groups. However, microadenomas were observed 10 weeks after the first treatment of DMH so that this protocol may be useful for studies of the early phase of colon carcinogenesis in mice.

Evaluation of apoptosis in human endometrial adenocarcinoma: comparison of nick end labeling and Le(y) antigen immunostaining method.

In situ estimation of DNA fragmentation by the nick end labeling method and immunohistochemical detection of cell surface carbohydrate Le(y) were performed to establish correlation between the results of these two apoptosis-detecting techniques in human solid tumor tissues. Formalin-fixed and paraffin-embedded tissue samples from 10 cases of human endometrial adenocarcinoma were examined and compared by using the two techniques. A substantial fraction of carcinoma cells was found to be definitely stained by both methods, although Le(y) expression did not seem to be positively correlated with DNA fragmentation in the carcinoma tissue. These results suggest that Le(y) expression does not necessarily reflect apoptotic DNA fragmentation, at least in human endometrial adenocarcinomas, and simultaneous application of DNA nick end labeling and Le(y) immunostaining is necessary to show the relevant signs of apoptosis in histologic slides, especially gynecological cancers.

Relationship between grade of intraductal component of breast cancer within main tumor and extent of intraductal spread in surrounding tissue.

Ninety-three specimens obtained by quadrantectomy of invasive ductal carcinoma of the breast were examined to evaluate the correlation between the grade of the intraductal component within the tumor and the extent of intraductal spread in the surrounding tissue. We used immunohistochemistry of a-smooth muscle actin to visualize the contour of intraductal components. The grade of the intraductal component within the tumor was classified into 4 groups (td0 to td3) by counting number of intraductal components; the extent of intraductal spread in the surrounding tissue was also divided into 4 groups (sd0 to sd3) according distance from the tumor margin. Fifty-eight (89.2%) of 65 cases with low-grade td were low-grade sd, and 17 (60.7%) cases of high-grade td were high-grade of sd. The grade of the intraductal component correlated with the extent of intraductal spread in the surrounding tissue, significantly (p < 0.001, Spearman rank correlation test). From these results, we believe that investigation of intraductal components within the tumor is useful for estimating the extent of intraductal spread in the surrounding tissue, and excision specimens from diagnostic lumpectomy should be examined for the extent of the intraductal component, the results being important in estimating the risk of local recurrence.

Occurrence of apoptotic DNA fragmentation in quiescent and proliferating cells in human endometrial adenocarcinoma tissues and the influence of apoptosis-suppressing effects of bcl-2 products.

In situ estimation of DNA fragmentation by the nick end labelling (NEL) method, and immunohistochemical examination of Ki-67 proliferative antigen and bcl-2 products in human endometrial adenocarcinoma tissues were performed to provide answers to the following two questions; a) does apoptotic DNA fragmentation occur specifically in quiescent cells or in proliferating cells or randomly in both?, b) does the bcl-2 product exert its apoptosis-suppressing effects differentially on carcinoma cells depending on their cell cycle condition?. Serial sections, one micrometer in thickness, from formalin-fixed and paraffinembedded tissues of 9 cases of human endometrial adenocarcinoma were examined. Apoptotic DNA fragmentation was observed in both quiescent (Ki-67 negative) and proliferating (Ki-67 positive) cells. Bcl-2 product-positive tumor cell islands tended to be NEL negative, although a few but non-negligible number of carcinoma cells, including both Ki-67 positive and negative ones, were NEL positive. These results indicate that, at least in human endometrial adenocarcinomas, apoptotic DNA fragmentation and bcl-2 product-independent (DNA) fragmentation occurs non-specifically with respect to the cell proliferation status. Further, the results suggest an altered regulation of cell death processes in human solid tumor tissue in vivo.

Inverse correlation between bcl-2 expression and cell growth fraction in human endometrial adenocarcinoma tissue.

Correlation between expression status of bcl-2 products and cell growth fraction (estimated by immunostaining for Ki-67 antigen) was analyzed in human endometrial adenocarcinoma tissue in situ. For this, serial sections, 2 micrometers thick, from formalin-fixed and paraffin-embedded tissue samples of 10 cases of the carcinoma were examined. Nine out of 10 carcinomas contained both bcl-2 positive and negative nests when examined immunohistochemically. In the remaining one case, no positive reaction for bcl-2 was observed. In general, bcl-2 positive nests tended to contain quiescent (Ki-67 negative) carcinoma cells, and bcl-2 negative nests, on the contrary, a large fraction of proliferating (Ki-67 positive) cells. However, this correlation was not strict, and in a few nests, the level of growth fraction was observed to be similar irrespective of bcl-2 positivity. These results show the complex function(s) of bcl-2 products, when considering their apoptosis-suppressing effects, cell cycle dependence and influence on cell proliferation status. Further, the results suggest an altered regulation of oncogene products in human solid tumor tissue in vivo.

Immunohistochemical detection of ras p21 oncoprotein in undifferentiated and well-differentiated epithelial carcinomas of the human ovary.

Expression of ras p21 oncoproteins in human ovarian carcinomas was examined immunohistochemically by using a monoclonal antibody(clone RAS 10) with respect to the degree of their histological differentiation. To achieve this, the intensity of staining for the protein was compared between undifferentiated and well differentiated carcinomas, i.e. extreme subtypes of common epithelial carcinomas. The former was composed of 8 "solid" carcinomas and the latter, 11 serous, 8 mucinous, 4 endometrioid and 4 clear cell carcinomas. All the cases examined, including both undifferentiated and well-differentiated carcinomas, showed a positive reaction to this antibody. Staining intensity and the number of positive cells somewhat varied among the cases. Additionally, 2 cases of ovarian epithelial tumors of low malignant potential (I,MP) were stained with this antibody. Both the cases were positive, but the number of positive cells seemed to be rather less than that found in the carcinoma groups. Thus, no differences in ras p21 expression were observed between the cases examined in spite of the differences in the degree of differentiation of the epithelial ovarian carcinomas. However, the possibility remained that the number of positive cells could be an indicator of malignant potential, enabling us to distinguish LMPs from carcinomas.

Tamoxifen mediated human endometrial carcinogenesis may not involve estrogenic pathways: a preliminary note.

Seven cases of endometrial adenocarcinoma patients who had experienced long-term tamoxifen treatments as adjuvant therapy of breast carcinoma, were investigated with respect to estrogen receptor (ER) status. Four cases of endometrial adenocarcinoma without tamoxifen treatment but with a previous history of breast carcinoma were investigated for comparison. One of the 7 and two of the 4 cases were positive for ER immunohistochemically. Thus, the frequency of ER positivity in secondary endometrial adenocarcinoma seemed to be at random among tamoxifen-treated and non-treated breast cancer patients. These results suggest that tamoxifen-mediated human endometrial carcinogenesis may not involve estrogenic pathway(s) but may involve other carcinogenic mechanisms such as DNA adduct formation as shown in rat liver tumorigenesis.