Randomized trial of preoperative docetaxel with or without capecitabine after 4 cycles of 5-fluorouracil­ epirubicin­cyclophosphamide (FEC) in early-stage breast cancer: exploratory analyses identify Ki67 as a predictive biomarker for response to neoadjuvant chemotherapy.

This randomized, multicenter study compared the efficacy of docetaxel with or without capecitabine following fluorouracil/epirubicin/cyclophosphamide (FEC) therapy in operable breast cancer and investigated the role of Ki67 as a predictive biomarker. Patients were randomized to 4 cycles of docetaxel/capecitabine (docetaxel: 75 mg/m2 on day 1; capecitabine: 1,650 mg/m2 on days 1­14 every 3 weeks) or docetaxel alone (75 mg/m2 on day 1 every 3 weeks) after completion of 4 cycles of FEC (5-fluorouracil 500 mg/m2, epirubicin 100 mg/m2 and cyclophosphamide 500 mg/m2 on day 1 every 3 weeks). The primary endpoint was the pathological complete response (pCR) rate. Predictive factor analysis was conducted using clinicopathological markers, including hormone receptors and Ki67 labeling index (Ki67LI). A total of 477 patients were randomized; the overall response in the docetaxel/capecitabine and docetaxel groups was 88.3 and 87.4 %, respectively. There were no significant differences in the pCR rate (docetaxel/capecitabine: 23 %; docetaxel: 24 %; p = 0.748), disease-free survival, or overall survival. However, patients with mid-range Ki67LI (10­20 %) showed a trend towards improved pCR rate with docetaxel/capecitabine compared to docetaxel alone. Furthermore, multivariate logistic regression analysis showed pre-treatment Ki67LI (odds ratio 1.031; 95 % CI 1.014­1.048; p = 0.0004) to be a significant predictor of pCR in this neoadjuvant treatment setting. Docetaxel/capecitabine (after 4 cycles of FEC) did not generate significant improvement in pCR compared to docetaxel alone. However, exploratory analyses suggested that assessment of pre-treatment Ki67LI may be a useful tool in the identification of responders to preoperative docetaxel/capecitabine in early-stage breast cancer.

Minimally invasive video-assisted thoracoscopic lobectomy for better clinical outcomes in peripheral T1NO lung cancer.

It is an unresolved issue whether various thoracotomies affect clinical outcomes. In addition, a wide variety of technical approaches of video-assisted thoracic surgery depend on the facility. We reviewed 152 consecutive patients with clinical T1N0M0 lung cancer that underwent three types of lobectomy with systematic mediastinal lymphadenectomy in a single institute: 46 conventional thoracotomies (OPEN), 50 anterolateral small thoracotomies mainly using the thoracoscope as a light guide (ASSIST), and 56 minimum thoracotomies in which only a thoracoscope view was used (PURE). Total discharge from the chest drainage tube, length of hospital stay, and post-thoracotomy pain were significantly less in PURE than in OPEN and ASSIST. The results of mediastinal lymphadenectomy were equivalent. The 3-year survival rates were also similar among the three groups. We conclude that good clinical outcomes, especially reduced post-thoracotomy pain, seemed to correlate with the lesser degree of destruction of the chest wall with the identical quality as an acceptable cancer operation in PURE.

Differential effects of estrogen and antiestrogen on in vitro clonogenic growth of human breast cancers in soft agar.

Of 534 human primary breast cancers provided for clonogenic assay in vitro, 276 (51.7%) developed distinctive colony formation by the soft-agar method. Estrogen receptors (ERs) were assayed by dextran-coated charcoal methods. A total of 65 (23.7%) of 274 breast cancers responded to added 10 nM 17 beta-estradiol (E2) by an increase in the number of colonies per dish of 150% or more of that in the controls treated with dextran-coated charcoal. The ER-positive and ER-negative tumors differed significantly in their response to E2: 55 (29.9%) of 184 ER-positive tumors responded versus 10 (11.1%) of 90 ER-negative tumors. The cancers in which the number of colonies increased to 150% or more of that of the controls were considered to be estrogen dependent; those in which the number of colonies increased to less than 150% of the control values were considered to be estrogen independent. When 1 microM tamoxifen (TMX) was added to the medium, 48 (21.3%) of 225 cancers showed a decreased in the number of colonies to 50% or less of that of the controls. Thus, we could separate breast cancers that were TMX sensitive (No. of colonies less than or equal to 50% of that of controls) from those that were TMX resistant (No. of colonies greater than 50% of that of controls). The response to TMX of the ER-positive cancers was significantly higher than that of the ER-negative tumors: 39 (25.5%) of 153 ER-positive tumors responded versus nine (12.5%) of 72 ER-negative tumors. In 153 ER-positive and 71 ER-negative tumors, we evaluated the correlation between the response to E2 and the response to TMX. ER-positive and ER-negative tumors differed significantly in their sensitivities to the two drugs. The TMX sensitivity did not completely correlate with the E2 dependence.(ABSTRACT TRUNCATED AT 250 WORDS)

In vitro clonogenic growth and metastatic potential of human operable breast cancer.

In 254 patients with operable [International Union against Cancer (1972) Stages I, II, and III] breast cancer, the relations between in vitro clonogenic growth in soft agar of primary breast cancer tumors and their metastatic potential expressed by the relapse-free survivals (RFS) as well as overall survivals were studied. Sixty-four % (163 of 254) of cancers formed distinct colonies (30 or more colonies in a single dish, or 10 or more colonies in plural dishes). Other breast cancers (36%, 91 of 254) were designated to be negative for the clonogenicity. There was no correlation between the positive or negative clonogenicity and clinicopathological characteristics in breast cancer patients, including the age of patients, menopausal status, tumor size, T classification, International Union against Cancer stage, histological type (Japanese Breast Cancer Society), histologically proved axillary lymph node metastasis, and estrogen receptor (ER). At the time of median follow-up of 43 mo (range, 25 to 61 mo) after mastectomy, a recurrence of malignancy occurred in 19.0% (31 of 163) of the patients with positive clonogenic tumors, and in 8.8% (8 of 91) of those with negative clonogenic tumors (P = 0.03). There also was a significant difference (P less than 0.03 by log rank test, P less than 0.05 by generalized Wilcoxon test) in RFS curves between positive and negative clonogenicity groups. These differences in RFS were also noted in Stage II patients in favor of the negative colony formation group. In ER-negative cancer patients, the RFS of patients with positive clonogenic cancers was shown to be worse (P less than 0.03 by log rank test, P less than 0.05 by generalized Wilcoxon test) than patients with negative clonogenic cancers. There was no difference in RFS in ER-positive cancer patients. There was a trend (P = 0.09 by log rank test) of worse overall survival rate in patients with positive clonogenic growth. In a multivariate comparison using the principal component analysis composed of factors including positive node, T classification, histological type, age, ER, and colony formation, the clonogenicity showed a significant effect on the recurrence of malignancy and also on the survival of the patients after mastectomy. In conclusion, in vitro clonogenic growth of the primary tumor of breast cancer was shown to be one of the independent factors of metastatic potential in operable breast cancer patients after mastectomy.

Update results of FEC followed by docetaxel neoadjuvant trials for primary breast cancer.

This study has been initiated to evaluate the safety, clinical and pathologic response as well as the relation of response (pCR or non-pCR) and survival (overall and relapse-free) of fluorouracil, epirubicin and cyclophosphamide (FEC) followed by docetaxel (DOC) as preoperative chemotherapy in patients with operable breast cancer. Japanese patients with primary breast cancer, Tlc-3N0M0 or T1-3NIM0, age 20-60, PS 0-1 were included in this study. Preoperative chemotherapy consisted of 4 cycles of FEC (500 mg/m(2), 100 mg/m(2), 500 mg/m(2)) every 3 weeks followed by 4 cycles of DOC (75 mg/m(2)) every 3 weeks. Since June 2002, 200 patients were enrolled in this study, and the time of this interim analysis, 80 patients were evaluable for safety and clinical efficacy. The overall clinical response rate was 71.4% (14% CR, 44% PR, 42% SD/PD), and the only G3,4 toxicities, neutropenia and febrile neutropenia were observed in 54% and 14% of patients, respectively. Eighty nine patients were evaluable for pathologic response by central review. Pathologic response was evaluated among invasive tumors on multiple cross-section specimens based on a modified version of the Japanese grading system for Japanese Breast Cancer Society. The pathologic response rate was 17%. In this ongoing trial, FEC followed by DOC was active and well tolerated.

Functions and ATP-binding responses of the twelve histidine residues in the TF1-ATPase beta subunit.

The C2 proton signals of all (twelve) histidine residues of the TF1 beta subunit in the 1H-NMR spectrum have been identified and assigned by means of pH change experiments and site-directed substitution of histidines by glutamines. pH and ligand titration experiments were carried out for these signals. Furthermore, the ATPase activity of the reconstituted alpha3beta3gamma complex was examined for the twelve mutant beta subunits. Two of three conserved histidines, namely, His-119 and 324, were found to be important for expression of the ATPase activity. The former fixes the N-terminal domain to the central domain. His-324 is involved in the formation of the interface essential for the alpha3beta3gamma complex assembly. The other conserved residue, His-363, showed a very low pK(a), suggesting that it is involved in the tertiary structure formation. On the binding of a nucleotide, only the signals of His-173, 179, 200, and 324 shifted. These histidines are located in the hinge region, and its proximity, of the beta subunit. This observation provided further support for the conformational change of the beta monomer from the open to the closed form on the binding of a nucleotide proposed by us [Yagi et al. (1999) Biophys. J. 77, 2175-2183]. This conformational change should be one of the essential driving forces in the rotation of the alpha3beta3gamma complex.

Pranlukast protects leukotriene C4- and D4-induced epithelial cell impairment of the nasal mucosa in vitro.

To investigate the effect of pranlukast on leukotriene- induced airway mucosal epithelial dysfunction, samples of human nasal mucosa obtained during surgery for facial trauma were exposed to leukotriene C4 and/or D4 and observed on a TV screen magnified x 2,500. Leukotriene C4- and D4-induced ciliary inhibition and delayed mucosal surface alterations appeared several hours later. Pranlukast prevented both the mucosal epithelial cell dysfunction and the delayed epithelial cell alteration.

Study on the metabolism of lyso-platelet activating factor (lyso-PAF) in human paranasal sinus mucosa. The cultured ciliated epithelium can convert lyso-PAF to PAF.

The conversion of lyso-platelet activating factor (lyso-PAF) to PAF in cultured paranasal sinus mucosa obtained from normal human subjects was studied. The PAF concentration in the medium was determined after addition of lyso-PAF. PAF became detectable at 10 minutes after the addition of 10(-8)M lyso-PAF, and reached a maximum concentration (3.25 x 10(-9)M) at 20 minutes. The PAF level then gradually declined to become undetectable at 60 minutes after addition of lyso-PAF. Thus PAF is very unstable having a half-life calculated to be 12.8 minutes with an elimination constant of k = 0.05377 minutes-1. In contrast, lyso-PAF is known to be a stable metabolite of PAF as well as a precursor of PAF. The results obtained from this study suggest that the turnover of lyso-PAF to PAF may play a role in evoking prolonged inflammation in target organs or tissues.

A possible direct action of a gonadotropin-releasing hormone agonist, goserelin, on the clonogenic growth of human breast cancer.

Direct inhibitory effects of a gonadotropin-releasing hormone agonist (GhRh agonist, goserelin, GOS) were studied on the in vitro clonogenic growth of human breast cancer, in comparison with estradiol-17 beta (E2) and tamoxifen (TAM). Estrogen (ER) and progesterone receptors (PgR) were asseyed in the adjacent cancer tissues using the DDC method. In 19% or 12 out of 64 cancers, E2, 10(-8) M, was shown to be effective in increasing the number of colonies per dish to 150% or more of the control. On the other hand, 16% (10/64) responded to TAM or GOS, respectively, i.e. the plating efficiency decreased to 50% or less of the control. While E2 and TAM had a tendency to have better effects in ER-positive tumors than ER-negative ones, the inhibitory effect of GOS was not dependent on the presence or absence of ER or PgR in the tumor. The GOS sensitivity did not correlate with the E2 dependence or the TAM sensitivity. These results suggest that a possible direct action of GOS may not be mediated by the ER system.

Steroid hormone receptors and response to high-dose medroxyprogesterone acetate in advanced breast cancer.

Forty-six patients with advanced breast cancer were treated with oral high-dose medroxyprogesterone acetate (MPA) as a second line endocrine treatment, with a 28% 13/46) response rate. There was a significant difference in response to MPA between patients with estrogen (ER)- or progesterone (PgR)-receptors and those with negative receptors assayed just before the treatment (ER+, 7/10, ER-, 0/9 of response rates, p = 0.007, PgR+, 4/4: PgR-, 3/15, p = 0.02). The combination of ER and PgR was observed to be best in predicting the response (p = 0.02). The ER status as well as the combination of ER and PgR, but not PgR alone detected in the primary tumors, correlated well with the response (ER+, 10/23: ER-, 0/15, p = 0.0094, PgR+: PgR-, p = 0.13, ER+PgR+, ER+PgR-, ER-PgR-:p = 0.024). The correlation between steroid hormone receptors and response to MPA in advanced breast cancer was established from the literature. Response rate to MPA was shown to be 42% (69/169) in ER+ cancer patients, and 10% (9/90) in ER- cancer patients (p = 0.00004). There was no correlation between PgR status and the response. Changes of receptors were studied in relation to the response. When ER+ or PgR+ tumors retained their positivity until MPA treatment, a good response was obtained, while there were almost no responders if one or both receptors changed from positive to negative, or remained negative. In conclusion, the response to MPA can be predicted by the steroid hormone receptors.

Age-related alterations in the auditory brainstem responses and the compound action potentials in guinea pigs.

The auditory brainstem response (ABR) and the eight nerve compound action potential (CAP) were measured using click click stimuli to investigate the age-related alteration in the auditory function in 66 guinea pigs consisting of four age groups. With advancing age, a gradual elevation of the thresholds in both the ABR and CAP was clearly seen, together with the prolonged latencies for waves I, II, III, and IV to clicks at 95 dBpeSPL in the ABR. There were some individual differences in either threshold elevation or latency prolongation of both the ABR and CAP in aged guinea pigs. These findings suggest that the effect of individual differences on degenerative aging processes of the auditory system should be considered in selected aged animals, although a significant elevation of the neural auditory threshold is clearly found with advancing age as a whole.

Platelet activating factor induced respiratory mucosal damage.

Human sinus mucosal specimens from eight normal individuals were exposed to platelet activating factor (PAF) at concentrations ranging from 10(-6) M to 10(-11) M in a humidified CO2 chamber at 37 degrees C. The mucosal surface of the specimens was recorded on video tapes and magnified 2,500 times on a 19-inch television (TV) monitor. Ciliary activity of each ciliated cell was photoelectrically measured on the TV monitor in real time. PAF induced mucosal damage which resulted in a coarse profile including ciliostasis and exfoliation of epithelial cells. The length of the incubation period in which the initial coarse profile occurred on the mucosal surface inversely correlated with the concentration of exposed PAF ranging from 10(-6) M to 10(-10) M with r = -0.712 (p less than 2 x 10(-4)). Both the control medium and 10(-8) M lysoPAF showed no effect on ciliary activity or mucosal surface alteration even after 24 hr of exposure. Significant ciliary inhibition was noted after 6 hr of exposure to PAF at concentrations of 10(-8) M and 10(-10) M (p less than 0.05). After 10 hr of exposure, significant ciliary inhibition (p less than 0.01) was noted at all concentrations. Inhibition occurred in a time- and dose-dependent manner. The length of the incubation period in which initial ciliostatis occurred and the level of PAF concentration showed an inverse correlation with r = -0.918 (p less than 10(-6)). These results indicate that PAF is cytotoxic to human respiratory mucosa.

Inhibition of mucociliary clearance of the eustachian tube by leukotrienes C4 and D4.

The effect of leukotrienes C4 (LTC4) and D4 (LTD4) and prostaglandin E2 (PGE2) on mucociliary clearance of the eustachian tube was investigated in vitro and in vivo. Normal ciliated epithelium was obtained from the eustachian tube of guinea pigs and incubated separately with LTC4, LTD4, and PGE2 at concentrations of 10(-8) mol/L and 10(-6) mol/L. Ciliary activity was measured photoelectrically. Leukotriene D4 progressively inhibited ciliary activity, while PGE2 promoted it. Leukotriene C4 also induced ciliary inhibition. One milliliter each of 10(-5) mol/L LTC4, LTD4, and PGE2 was directly injected into the tympanic bullae of chinchillas under anesthesia. The middle ears were examined by otomicroscopy, tympanometry, and auditory brain stem response over time. Clearance of middle ear effusion was delayed by LTC4 and LTD4, as compared with PGE2 and the control. These findings indicate that LTC4 and LTD4 inhibit mucociliary clearance of the eustachian tube.

Interaction between oral alpha-streptococci and group A streptococci in patients with tonsillitis.

The incidence of oral alpha-streptococci with inhibitory activity against group A streptococci, as a defense mechanism against bacterial infection in the oral cavity, was investigated in 141 patients with streptococcal tonsillitis. The study population included both children (n = 79) and adults (n = 62). Infection by group A streptococci appeared to be more common in children than in adults, as the detection rates of inhibitory alpha-streptococci in healthy children (29.7%), as well as pediatric patients with tonsillitis (14.9%), were lower than those in adults (63.0%; p < .01). It is possible to consider oral alpha-streptococci with inhibitory activity to be among the indications for tonsillectomy in patients with streptococcal tonsillitis, since the detection rate of inhibitory alpha-streptococci in surgical cases (10.9%) was significantly lower than that in nonsurgical cases (31.1%; p < .01). The high detection rate of these strains during the postoperative state supported the observation that the incidence of group A streptococcal infection was decreased postoperatively. Accordingly, it is useful to investigate bacterial interference between oral alpha-streptococci and group A streptococci in patients scheduled for tonsillectomy.

Effects of platelet activating factor on mucociliary clearance of the eustachian tube.

The effects of platelet activating factor (PAF) on mucociliary clearance of the eustachian tube were investigated both in vitro and in vivo. Normal ciliated epithelium was obtained from the eustachian tube of guinea pigs and incubated with PAF at concentrations ranging from 10(-10) to 10(-6) mol/L. Ciliary activity was observed under an inverted microscope and quantified photoelectrically. The PAF dose-dependently inhibited ciliary activity. One milliliter each of 10(-5) mol/LPAF, 10(-5) mol/L prostaglandin E2 (PGE2), 10(-5) mol/LPAF and PGE2, or the control solution (0.1 v/v% methanol-phosphate-buffered saline) was directly injected into the tympanic bullae of anesthetized chinchillas. The middle ear was examined by otomicroscopy, tympanometry, and auditory brain stem response in relation to time. The PAF delayed middle ear clearance, and the PGE2 augmented its delay. These findings suggest that PAF inhibits mucociliary clearance of the eustachian tube from the middle ear, and that PGE2 plays an important role in the augmentation of inflammatory disorders.

Peripheral arterial coil embolization for hepatic arteriovenous malformation in Osler-Weber-Rendu disease; useful for controlling high output heart failure, but harmful to the liver.

A 55-year-old Japanese housewife, who had Osler-Weber-Rendu disease, was admitted to our hospital because of frequent epistaxis and worsening exertional dyspnea. The computed tomography and hepatic arteriography revealed large hepatic arteriovenous malformation, which was considered to be the leading cause of her high output heart failure. Two series of hepatic arterial coil embolization procedures were performed to reduce hepatic shunt flow. They temporarily improved her cardiac condition, but gradually induced progressive hepatic failure due to intrahepatic cholangitis. Hepatic dysfunction restricted her quality of life and lead to a fatal clinical course one year after the second coil embolization.

Investigation of oral alpha-streptococcus showing inhibitory activity against pathogens in children with tonsillitis.

The incidence of oral alpha-streptococcus with inhibitory activity against group A streptococcus, as a defense mechanism against bacterial infection in the oral cavity, was investigated in pediatric individuals with tonsillitis. Infection by group A streptococcus appeared to be common in children, because the detection rate of inhibitory alpha-streptococcus in healthy children as well as pediatric patients with tonsillitis was lower than in adults and elderly patients. In particular, the detection rate of these strains was predominantly low in patients with beta-streptococcus. Among pediatric patients scheduled for tonsillectomy, the detection rate of inhibitory alpha-streptococcus was low preoperatively. However, the rate was markedly increased after surgery. The high postoperative detection rate of these strains reflected the decreased incidence of group A streptococcal infection. The results of this investigation of bacterial interference between oral alpha-streptococcus and group A streptococcus suggested that surgical treatment is a more effective approach for improving the oral bacterial flora in children with recurrent tonsillitis.

Mucosal dysfunction and damage induced by platelet activating factor (PAF).

The effect of PAF on human nasal mucosa was investigated in vitro. Normal paranasal sinus mucosa was obtained from the ethmoid sinuses by surgical procedure and incubated in the form of tissue culture. Ciliary movement was viewed under an inverted microscope and recorded on video tapes, and its activity was measured photoelectrically. Morphological alterations were examined by light and electron microscopy. PAF inhibited ciliary activity of human nasal mucosa, in a time and a dose dependent manner, at concentrations from 10(-6)M to 10(-10)M, while no significant change was observed at 10(-11) M. Lyso-PAF exhibited minimal effect on the mucosa at a concentration of 10(-6) M. Morphological alterations of the epithelial layer of the mucosa such as edema, cell exfoliation and desquamation were found to increase across time. Ultrastructural alterations were observed prior to inhibition of ciliary activity. These data indicate the cytotoxic effect of PAF on human paranasal sinus mucosa.

Promotive effect of lysozyme on the ciliary activity of the human nasal mucosa.

Lysozyme is an important resistance factor in the respiratory defence mechanism. Few reports exist concerning its effects on the ciliary activity of the human nasal mucosa. To clarify the biological activity of lysozyme in ciliary movement, a quantitative study was undertaken using a photo-electrical method to measure ciliary beats in vitro under conditions of constant temperature. Egg-white lysozyme in a 0.01 M sodium phosphate buffer solution, pH 7.4, dose-dependently accelerated the ciliary beats in the human nasal mucosa, removing overlying mucus continuously for at least 60 min. The buffer alone, however, failed to affect ciliary beats. These results indicate that egg-white lysozyme directly promotes the ciliary beats in the human nasal mucosa.

Efficacy of emulsion containing Gd-DTPA and lipiodol in hepatic transcatheter arterial embolization.

We evaluated the clinical efficacy of an embolizing emulsion produced by mixing lipiodol and Gd-DTPA, in transcatheter arterial chemoembolization (TAE). Subjects were 10 patients with hepatocellular carcinoma (HCC). The emulsion used was produced by mixing 3 ml of lipiodol and anticancer agents (mitomycin-C 10 mg and adriamycin 20 mg) dissolved in Gd-DTPA. This emulsion was infused into the proper hepatic artery. Subsequent embolization by Gelfoam was performed in eight patients. MRI and CT examinations were performed soon after TAE (1 or 2 days after) and two weeks afterwards. The position of lipiodol accumulation dipicted on CT at two weeks after TAE did not differ from the site of change in signal intensity induced by Gd-DTPA on MR images soon after TAE in any case. In almost all cases, the washout of Gd-DTPA occurred earlier than that of lipiodol. It might be suggested that Gd-DTPA, which is water-soluble, shows in vivo dynamics similar to anticancer agents rather than to lipiodol, which is oil-soluble. Since the normal tissues showed no definite signal changes, we could easily detect the site of tumors by using the emulsion containing Gd-DTPA even on MR studies immediately after TAE. In addition, the deposits of Gd-DTPA depicted on MR images created fewer artifacts than the lipiodol deposits on CT.