Periacetabular osteotomy reduces the dynamic instability of dysplastic hips.

We compared the dynamic instability of 25 dysplastic hips in 25 patients using triaxial accelerometry before and one year after periacetabular osteotomy. We also evaluated the hips clinically using the Harris hip score and assessed acetabular orientation by radiography before surgery and after one year. The mean overall magnitude of acceleration was significantly reduced from 2.30 m/s(2) (sd 0.57) before operation to 1.55 m/s(2) (sd 0.31) afterwards. The mean Harris hip score improved from 78.08 (47 to 96) to 95.36 points (88 to 100). The radiographic parameters all showed significant improvements. This study suggests that periacetabular osteotomy provides pain relief, improves acetabular cover and reduces the dynamic instability in patients with dysplastic hips.

Comparison of activities of enzymes related to energy metabolism in peripheral leukocytes and livers between Holstein dairy cows and ICR mice.

Activities of enzymes related to energy metabolism and isoenzyme patterns of lactate dehydrogenase (LDH) were determined in peripheral leukocytes and livers of Holstein dairy cows and Institute of Cancer Research (ICR) mice. In dairy cow liver, activities of enzymes in glycolysis, malate-aspartate shuttle and lipogenesis were lower, but activities of glucose-6-phosphatase in gluconeogenesis were higher than those in mouse liver. Glucokinase activities were below detection limit in leukocytes and liver of the cows. Dairy cow leukocytes and liver showed the isoenzyme patterns with dominance of LDH-1, -2 and-3, whereas mouse leukocytes and liver showed that LDH-5 was dominant. The LDH isoenzyme patterns were very similar between leukocytes and liver in each animal species. Some enzymes in leukocytes may reflect those enzymes activities in liver and be a useful indicator for energy metabolism in animals.

Application of Cre-loxP system to the urinary tract and cancer gene therapy.

The bacteriophage P1-derived Cre-loxP system is a powerful and versatile tool for in vivo DNA recombination. It is widely used in gene targeting research. The combination of the Cre-loxP system and a specific promoter enables conditional "knockout" of a target gene in a particular tissue or cell type. It has also made it possible to delete genes in adult animals that are essential for embryogenesis. This system is also used to enhance tissue- or tumor-specific promoter activities that are useful for gene therapy of certain types of cancer. It is expected that this simple and effective system will be further utilized in various research applications.

Suicide gene therapy on LNCaP human prostate cancer cells.

The efficacy of combination suicide gene therapy was evaluated using a Herpes simplex virus-thymidine kinase/ganciclovir (HSV-TK/GCV) system and an Escherichia coli cytosine deaminase/5-fluorocytosine (CD/5-FC) system on the LNCaP human prostate cancer cell model. Two types of plasmid vectors with the HSV-TK gene were constructed. A constitutive chicken beta-actin promoter drove one and a prostate-specific antigen (PSA) promoter drove the other. Similarly, a pair of plasmids with the CD gene under a cytomegalovirus (CMV) promoter and the PSA promoter was also constructed. LNCaP cells were transfected in vitro with either or both of those plasmids using a cationic lipid reagent. Transfected cells were treated with GCV and/or 5-FC. The percentage of viable LNCaP cells 7 days after treatment with HSV-TK/GCV or CD/5-FC under a constitutive promoter was 40% and 41% of controls, respectively. The cell viability when two suicide genes were combined was 23%. The cell viabilities after four days with PSA promoter-HSV-TK vectors, CD vectors and a combination of both were 79%, 88% and 88%, respectively. Suicide gene therapy using either HSV-TK/GCV, CD/5-FC, or both, was effective in the LNCaP model. An additive effect was observed when the two suicide genes were used together. The PSA promoter did not seem to be effective enough to elicit cytotoxicity under the experimental conditions used here.

[Immunohistochemical study of p53 and Ki-67 overexpression in grade 3 superficial bladder tumor in relationship to tumor recurrence and prognosis].

PURPOSE: The major drawback of the current treatment for superficial bladder tumor is the high rate of recurrence. Especially, the tumor with grade 3 component has a tendency to recur and progress in stage. However, we have difficulty in predicting tumor recurrence and stage progression accurately by conventional clinicopathological factors. We evaluated the efficacy of p53 and Ki-67 overexpression as a predictor of recurrence or prognosis in patients with superficial bladder tumor of grade 3. MATERIALS AND METHODS: Samples were obtained from 41 patients with superficial transitional cell carcinoma of the bladder of grade 3 who were treated by transurethral resection (TUR). The immunohistochemical study was performed using the antibodies against the p53 protein and Ki-67 antigen on formalin-fixed, paraffinembedded tissue specimens from initial tumors. We evaluated the correlation between these results and several clinicopathological factors. RESULTS: The p53 index and the Ki-67 index in pTa, pT1a and pT1b tumors were 26.4 +/- 30.1%, 28.6 +/- 30.0%, and 34.6 +/- 32.6% (p53) and 20.5 +/- 22.5%, 20.0 +/- 29.3%, and 29.2 +/- 28.4% (Ki-67). There was no significant difference between the each index and tumor stage. Eighteen cases (43.9%) had intravesical recurrence. The p53 index of the initial tumor from the tumor free cases (n = 23), recurrent cases without stage progression (n = 12), and stage progression cases (n = 6) were 19.7 +/- 28.2%, 42.0 +/- 28.7%, and 42.5 +/- 32.0%. Between the recurrence-free cases and the recurrent cases without progression, the p53 index of the initial tumor had statistical significance (p < 0.05). The Ki-67 index was shown to be the same pattern as the p53 index, but there was not statistical significance. Four of patients with stage progression had tumor progression within six months. Three of the patients with tumors with stage progression died of the cancer. In multivariate analysis, tumor multiplicity (p = 0.01), BCG intravesical instillation (p = 0.04), p53 index (p = 0.01) and Ki-67 index (p = 0.02) were the positive risk factors for tumor recurrence, but only the p53 index was the positive risk factor for prognosis fo the patients (p = 0.03). CONCLUSION: These results suggest that the immunohistochemical study of p53 overexpression is a useful predictor for tumor recurrence and prognosis in patients with superficial bladder tumor with grade 3.

Os vesalianum as a cause of lateral foot pain: a familial case and its treatment.

This is a rare case of bilateral symptomatic os vesalianum in a 13-year-old girl whose mother had the same condition unilaterally. We performed osteosynthesis and bone grafting instead of a simple resection to preserve the peroneus brevis tendon, with excellent results.

An Interlaboratory Validation Study of the Improved Transformation Assay Employing Balb/c 3T3 Cells: Results of a Collaborative Study on the Two-stage Cell Transformation Assay by the Non-genotoxic Carcinogen Study Group.

The Non-genotoxic Carcinogen Study Group of the Environmental Mutagen Society of Japan organised the first step of an interlaboratory validation study on an improved cell transformation assay employing Balb/c 3T3 A31-1-1 cells. Nineteen laboratories participated in this study. The modified transformation assay was evaluated for its responsiveness, its interlaboratory reproducibility and its transferability. In this study, a mixture of Dulbecco's modified Eagle's medium and nutrient mixture F12, supplemented with insulin-transferrin-ethanolamine-sodium selenite and 2% fetal bovine serum (FBS) was used during the period of expression of transformed foci, intead of the usual minimum essential medium with 10% FBS. 20-Methylcholanthrene (MCA) and 12-O-tetradecanoylphorbol-13-acetate (TPA) were selected as a prototype initiator and a tumour promoter, respectively. Two series of experiments were conducted. In the first series, the transformation activity of MCA was examined at various concentrations. In the absence of the promoting treatment with TPA, exposure to MCA only weakly induced transformed foci. In the presence of 0.1µg/ml TPA, all laboratories observed significant dose-dependent increases in the number of transformed foci with increasing MCA concentrations. In the second series of experiments, various concentrations of TPA were tested. In the absence of initiating treatment with MCA, exposure to TPA weakly induced transformed foci in about half of the laboratories. In the presence of 0.2µg/ml MCA, all the laboratories observed significant dose-dependent increases in the number of transformed foci with increasing TPA concentrations. The results from this study support the usefulness of this modified two-stage transformation assay with Balb/c 3T3 cells.

Tree-type algorithm for statistical analysis in chronic toxicity studies.

An appropriate statistical methodology in toxicity studies has been discussed over the last two decades and many statistical methods have already been proposed. Many practical problems, however, still remain unresolved and most pharmaceutical industries have been using a tree-type algorithm routinely to analyze repeated-dose toxicity study data. In considering routine use of statistical analysis in toxicological studies, standardization of statistical methodology is necessary and the decision tree has an important role. In this article, the problems, relating to tree-type algorithms are summarized. Then we propose a new tree-type algorithm, which targets quantitative data in repeated-dose studies in rodents, usually sample size per group between 10 to 20, based on the following two important principles: "using a parametric method" and "suitable for intuition of toxicologists". An example of its application to actual toxicity study data is demonstrated. The performance of this new method is also evaluated using historical data. However, it should be noted that the intention of this paper is not to make a definite solution of the decision tree. Several other alternatives can be considered. Since there is no single theoretically correct solution of tree-type algorithms, too formal a use of the decision tree is not recommended. We must not forget the exploratory nature of evaluating repeated toxicity data.

Analysis of synergism in hepatocarcinogenesis based on preneoplastic foci induction by 10 heterocyclic amines in the rat.

The effects of simultaneous treatment with 5 or 10 heterocyclic amines at low dose levels on hepatocarcinogenesis in rats were investigated using a medium-term liver bioassay protocol based on the two-stage carcinogenesis hypothesis with diethylnitrosamine initiation (200 mg/kg, i.p.). Five carcinogenic heterocyclic amines in experiment 1 (Trp-P-1, Glu-P-2, IQ, MeIQ, MeIQx) and experiment 2 (Trp-P-2, Glu-P-1, MeAalphaC, AalphaC, PhIP) were administered together or individually in the diet at levels of 1/1, 1/5, or 1/25 carcinogenic doses, and all 10 chemicals were given at 1/10 or 1/100 levels in experiment 3. Induction of preneoplastic glutathione S-transferase placental form (GST-P)-positive foci in the liver was generally increased in the combination groups over the sums of the 5 or 10 individual effects. Thus, based on the heteroadditive concept, synergism was observed for each combination, being most obvious in the group given all 10 chemicals at the 1/10 dose levels. However, the values for the combined groups were generally close to the averages of the 5 or 10 data gained for the heterocyclic amines alone at the corresponding higher doses, indicating the possibility of isoadditivity. Based on these findings, we propose here a new statistical method for analysis of combined effects of multiple chemicals, and, using this, we demonstrated (true) synergism with some heterocyclic amine combinations. The importance of dose-response curves for evaluation of combination effects is discussed.

[Whole mouth gustatory test (Part 2)--Effect of aging, gender and smoking on the taste threshold].

The whole mouth test method was employed for examining changes in gustatory senses due to aging. Participants were 314 male and 356 female healthy volunteers not complaining of taste disorders. Simultaneously, the effects of smoking were examined, comparing 71 male smokers to 142 male non-smokers, ranging from 20 to more than 60 years of age. A multiple comparison procedure (Tukey-Kramer) was applied to age groups, revealing a significant increase in the threshold with age; high threshold values were observed in the "70s and older" group in comparison to the younger group for all tastes other than the sweet taste. Concerning the differences between males and females, the gustatory thresholds of males were found to be higher than those of females for some in the "20s and older" group and for all tastes other than the sweet taste, whereas the thresholds of males were lower than those of females for all taste qualities in the late teen group. As to male subjects in whom the effects of smoking could be studied, smokers in their 20s demonstrated a slight rise in the threshold only for the bitter taste.

[Influence of aging on electrogustometric threshold].

Electrogustometric testing was carried out, with one researcher examining the electrogustometric threshold at three sites, in 461 normal cases, by TR-06 (Rion Co.), over a wide range. A multiple comparison procedure (Tukey-Kramer) was applied to age groups, revealing a significant increase in the threshold with advancing age; high threshold values were observed in the "60s and older" group for the chorda tympani nerve area (tongue tip) and the glossopharyngeal nerve area (tongue base), and in the "70s and older" group for the superficial greater petrosal nerve area (soft palate). As to the gender difference in women, a significant drop was observed in the late teens whereas a low tendency in all sites tested was not statistically, significant in other age groups. Differences between the right and the left sides were seen in most cases but were within 6dB. As to site differences, the soft palate threshold was higher than those of the tongue tip and the tongue base in all age groups. A significant decrease in the threshold of the tongue tip, as compared to the tongue base, was observed for those in their late teens, whereas minimal differences were noted in the other age groups and at other sites. The soft palate threshold was significantly reduced in smokers in their 30s and 40s as compared with non-smokers.

p53 gene mutation in recurrent superficial bladder cancer.

Surgical specimens from 30 patients with transitional cell carcinoma of the bladder were examined for the presence of mutation of the p53 tumor suppressor gene using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis. All of the specimens were superficial, low grade cancer, and 10 patients had recurrences during an observation period of 24 months following the initial surgical treatment. Among the 30 patients with superficial bladder cancer, 4 were found to have the mutated p53 gene and 3 of them had recurrences, which involved either invasive or high grade tumors. Two of these recurrent tumors with the p53 gene mutation showed identical mutations with those primary tumors, whereas the other one did not have the same mutation. The data suggest that bladder cancers with a mutation of the p53 gene have a greater probability of poor prognosis than those which do not, even if the primary lesion was a superficial, low grade tumor. Therefore, the presence of the p53 gene mutation may provide important clues to the factors involved in bladder cancer.

Effects of 5 alpha-dihydrotestosterone (DHT) on the transcription of nm23 and c-myc genes in human prostatic LNCaP cells.

Proliferation of the androgen-dependent human prostate LNCaP cells was increased by the androgen DHT. Changes in the steady state level of the nm23 and c-myc mRNA in LNCaP cells, with or without 10 nM DHT, showed the nm23 mRNA to change rapidly, beginning to rise after 2 h and reaching its peak by 4 h of DHT treatment. In contrast, increases in the c-myc gene only became apparent after 4 h of treatment. Maximal increase of nm23 mRNA was observed at 10(-9) M DHT. The basal expression of c-myc and nm23 mRNAs was between 30-70% lower in the LNCaP cells, as compared with the androgen-independent PC-3 and JCA-1 human prostatic human carcinoma cells. Thus nm23 may be classified as a member of the early androgen-responsive genes.

[Tachistoscope and dichotic listening test of the subject after the transection of the posterior part of the corpus callosum].

The authors made neuropsychological studies by the tachistoscope and the dichotic listening test on a subject who had undergone the transection of the posterior part of the corpus callosum. As to the tachistoscopic recognition, stimulus material was composed with the various Japanese letters (Katakana, Hiragana, Kanji), various faces (variations of the eyebrow form and the mouth form) and various slopes of line. Table 1 shows results of the cases (the subject was the present case, subjects 1 and subject 2 were past cases). It was seen that the performance of the subject on Japanese letters tasks showed greater right visual field superiority than the one of subject 1 and subject 2. As to the auditory recognition, the tasks used for the dichotic listening test were the following (Table 2, 3, 4). Different digits (three pairs) of the subject showed greater right ear superiority (right ear: 61.1, left ear 5.9) than the ones of subject 1 and subject 2.

[Neuropsychological studies of two subjects after the transection of the splenium of the corpus callosum (Part 3. The tactile recognition) (author's transl)].

We made the neuropsychological tactile studies of two right-handed subjects who had undergone the transection of the splenium of the corpus callosum for the pineal operation (the teratoma by the histopathological examination). The transaction grade was 2.0 cm and 2.5 cm. In this paper we examined the tactile recognition neuropsychologically: A. Superficial sensory modalities: light touch, pain perception, temperature discriminations. B. Deep sensory modalities: vibration perception, position sense, motor sense. C. Perception of material: rough or smooth, heavy or light, hard or soft. D. Perception of size: appreciation of shape in two dimentions: circle, triangle, square, cross, rhomb, appreciation of form in three dimensions: ball, column, cone, trignonal prism, cube. E. Identification of simple objects: eraser, pencil, hair pencil, fountainpen, telephone, glasses. F. Identification of japanese letters: Kana (phonetic symbols) and Kanji (essentially nonphonetic logographic symbols representing lexical morphemes). According to the papers of Akelaitis, Van Vleuten, Goldstein, the subjects after the transection of the corpus callosum could name the objects and the card board letters placed in their right hands with their eyes closed, but they could not name in their left hand by contrast. In our subjects, the identification of simple objects and the identification of japanese letters were complete good. But they had the neuropsychological visual and auditive disorders. The result shows that the commissure fibres of the identification of the simple objects and letters of two hemispheres does not connect through the splenium of the corpus callosum.

[Neuropsychological studies of two subjects after the transection of the splenium of the corpus callosum. (Part 2. The auditory recognition by the dichotic listening test) (author's transl)].

We made the neuropsychological auditory studies of two right-handed subjects who had undergone the transection of the splenium of the corpus callosum for the pineal operation (the teratoma by the histopathological examination). The transection grades were 2.0 cm and 2.5 cm. The truncus and splenium of the second subject were flat by the compression of the tumor. We examined these subjects by the some audiolocigal methods. 1. Pure tone audiometry. 2. Fixed frequency Békésy audiometry. 3. SISI test (short increment sensitivity index test). 4. Speech audiometry. 5. Distorted speech test (filtered speech test). 6. Binaural fusion test. 7. Dichotic listening test. The tasks used for the dichotic listening test were following. Different digits (one pair, three pairs), different words (from two to four syllables) and different vowels and consonants were presented simultaneously to the two ears by means of a dual-channel tape recorder with stereophonic earphone. The result shows that the first subject (whose section of the spelenium was 2.0 cm) shows a slight but not significant superiority on the right ear to the one pair digits and words (p less than 0.1), and the second subject (whose section of the splenium was 2.5 cm and whose splenium and truncus of the corpus callusum were flatten by the compression of the tumor) shows a significant superiority on the right ear to the one pair digits, words, three pairs digits and vowels (p less than 0.05). But there was no definitive abnormalities at the pure tone audiometry, the fixed frequency Békésy audiometry, SISI test, distorted speech test and binaural fusion test of two cases. The result shows that the commissure fibres of the auditory perception of the speech of two hemispheres connect through from splenium to the truncus of the corpus callosum.