The Impact of Pathological Grade Group 3 on Relapse-free Survival After Salvage Radiotherapy for Postoperative Prostate Cancer.

BACKGROUND/AIM: This retrospective study aimed to investigate the outcomes of relapse-free survival (RFS) after salvage radiation therapy (SRT) to the prostate bed for postoperative biochemical recurrence of prostate cancer. PATIENTS AND METHODS: A total of 87 patients were analyzed. There were 27, 32, and 24 patients with pathological grade groups of 1-2, 3, and 4-5, respectively. SRT doses of 64, 66 or 70 Gy were administered to 24, 3 and 60 patients, respectively. The Kaplan-Meier method was used to estimate time-to-event outcomes. The multiple imputations method was used to impute missing values, and Cox proportional-hazards models were applied for multivariate analyses. RESULTS: The median follow-up period for patients overall was 58.6 months. The 5-year RFS rates of the whole cohort was 59.4% and those for pathological grade groups 1-2, 3 and 4-5 were 88.9%, 37.7% and 39.5%, respectively. In multivariate analyses, higher pathological grade group [4-5 vs. 3 vs. 1-2: hazard radio (HR)=8.65, p<0.01], negative surgical resection margin (positive vs. negative: HR=0.41, p=0.02) and higher pre-salvage treatment serum prostate-specific antigen (cutoff value 0.31 ng/ml: HR=3.50, p<0.01) were significantly associated with poorer RFS. The cumulative incidences of grade 2 or more late rectal bleeding and late hematuria were 4.9% and 8.7%, respectively, at 5 years and 4.9% and 15.7%, respectively, at 8 years. These toxicities occurred only in the 70 Gy-treated arm. CONCLUSION: Our study revealed that pathological grade group 3 prostate cancer patients experienced moderately unfavorable RFS after SRT. Higher radiation doses might increase late toxicities without improving RFS.

Recent Postoperative Radiotherapy for Left-sided Breast Cancer Does Not Increase Mortality of Heart Disease in Asians or Pacific Islanders: SEER Database Analysis.

BACKGROUND/AIM: The purpose of this study was to evaluate the impact of recent radiotherapy on mortality from heart disease in Asians or Pacific islanders with breast cancer using the Surveillance, Epidemiology, and End Results (SEER) database. PATIENTS AND METHODS: Asians or Pacific islanders with stage 0 or I (AJCC 6th) breast cancer between 2000 and 2015 were analyzed. The impact of radiotherapy for mortality from heart disease after treatment was evaluated by comparing patients who received radiotherapy for left-sided breast cancer, patients who received radiotherapy for right-sided breast cancer and patients who did not receive radiotherapy. RESULTS: In 25,684 Asians or Pacific islanders, the incidence of cardiac death was higher in patients who did not receive radiotherapy than in patients who received radiotherapy. Among patients who received external beam irradiation, the incidence of cardiac death was 2.00% for patients with left-sided breast cancer and 1.69% for patients with right-sided breast cancer, with no significant difference (chi-square test, p=0.427). In the period from 2000 to 2008, there was no significant difference between the cumulative heart-related death rates in patients who received radiotherapy and in patients who did not receive radiotherapy (Tarone-Ware test, p=0.406); however, in 2009-2015, the cumulative heart-related death rate in patients who did not receive radiotherapy was significantly higher than that in patients who received radiotherapy (log-rank test, p<0.001). CONCLUSION: Heart-related death after treatment for breast cancer is relatively rare in Asians or Pacific islanders. Since at least 2000, the cardiac impact of postoperative radiotherapy has not been significant.

Delivery of RANKL-Binding Peptide OP3-4 Promotes BMP-2-Induced Maxillary Bone Regeneration.

Although bone morphogenetic protein 2 (BMP-2) is known to stimulate osteogenesis, there is evidence that high doses of BMP-2 can lead to side effects, including inflammation and carcinogenesis. The supplementation of other bone-augmenting agents is considered helpful in preventing such side effects by reducing the amount of BMP-2 required to obtain a sufficient amount of bone. We recently showed that a receptor activator of nuclear factor κB ligand (RANKL)-binding peptide promotes osteoblast differentiation. In the present study, we aimed to investigate whether OP3-4, a RANKL-binding peptide, promotes BMP-2-induced bone formation in the murine maxilla using an injectable gelatin hydrogel (GH) carrier. A GH carrier containing OP3-4 with BMP-2 was subperiosteally injected into the murine maxillary right diastema between the incisor and the first molar. The mice were sacrificed 28 d after the injections. The local bone formation in the OP3-4-BMP-2-injected group was analyzed in comparison to the carrier-injected, BMP-2-injected, and control-peptide-BMP-2-injected groups. The GH carrier containing OP3-4 with BMP-2 enlarged the radio-opaque area and increased the bone mineral content and density in the radiological analyses in comparison to the other experimental groups. Interestingly, fluorescence-based histological analyses revealed that the mineralization had started from the outside, then proceeded inward, suggesting that the size of the newly formed bone had already been set before calcification started and that the effects of OP3-4 might be involved in accelerating the early steps of osteogenesis. Actually, OP3-4 enhanced the BMP-2-induced 5-bromo-2'-deoxyuridine (BrdU)-positive cell numbers at the injected site on day 7 and the expression of Runx2 and Col1a1, which are early osteogenic cell markers, on day 10 after the subperiosteal injections. In summary, we demonstrated, for the first time, that the application of OP3-4 by subperiosteal injection promoted BMP-2-induced bone formation, which could lead to the development of an easy and noninvasive means of promoting alveolar ridge formation.

Relationship between perceived softness of bilayered skin models and their mechanical properties measured with a dual-sensor probe.

The development of a sensor system that can predict the subjective softness of human skin is an important goal for the cosmetics industry. Here, we first carried out a subjective softness evaluation test using 65 skin models consisting of polyurethane bilayers with different thickness of the superficial layer and different degree of cross-polymerization of the basal layer. The results showed that perceived softness was dependent on the mechanical properties of both the superficial and basal layers. Then, we used a recently developed tactile sensor system composed of a piezoelectric tactile sensor and a load cell to measure mechanical softness parameters of the superficial layer and the whole model, respectively. Statistical analysis showed that the data obtained from these two sensors were well correlated with the perceived softness of the prepared models. These results suggest that it may be feasible to predict the subjective softness of human skin in vivo from non-invasive mechanical softness measurements of the superficial skin layer and whole skin obtained with our new dual-probe sensor system.

Tactile resonance sensors in medicine.

Tactile sensors in general are used for measuring the physical parameters associated with contact between sensor and object. Tactile resonance sensors in particular are based on the principle of measuring the frequency shift, Deltaf, defined as the difference between a freely vibrating sensor resonance frequency and the resonance frequency measured when the sensor makes contact to an object. Deltaf is therefore related to the acoustic impedance of the object and can be used to characterize its material properties. In medicine, tactile resonance sensor systems have been developed for the detection of cancer, human ovum fertility, eye pressure and oedema. In 1992 a Japanese research group published a paper presenting a unique phase shift circuit to facilitate resonance measurements. In this review we summarize the current state-of-the-art of tactile resonance sensors in medicine based on the phase shift circuit and discuss the relevance of the measured parameters for clinical diagnosis. Future trends and applications enabled by this technology are also predicted.

A new tactile skin sensor for measuring skin hardness in patients with systemic sclerosis and autoimmune Raynaud's phenomenon.

We used a new tactile sensor to measure the elastic properties of skin in patients with systemic sclerosis or Raynaud's phenomenon. The sensor consists of a piezoelectric vibrator with vibration pickup to measure frequency changes when the sensor is placed on the skin. The mean frequency change at the skin surface of the proximol third phalanx in patients with systemic sclerosis was significantly lower than in age- and sex-matched controls. The results in systemic sclerosis patients were statistically correlated to the Modified Rodnan Skin Thickness Score. This technique was also used to measure the therapeutic efficacy of salpogrelate, a new specific serotonin receptor antagonist. A greater mean frequency change was seen after treatment. We conclude that this new tactile sensor is useful for quantitatively measuring skin sclerosis and may help determine the efficacy of therapeutic treatments.

Development of a vibratory microinjection method.

To reduce cellular damage by pronuclear microinjection and nuclear transfer, we have recently developed a vibratory microinjection method. A micropipette was fixed to a piezoelectric ceramic with a resonance frequency of 70 kHz. When this micropipette was vibrated, it easily entered a mouse-fertilized egg without any sharp depression of the cell body, whereas a sharp, deep depression at the insertion site was observed when the micropipette was not vibrated. A depression rate defined as a rate of a depth of depression over an original cell diameter was utilized as an index of cellular deformation. The depression rates with and without vibration were 11.1 +/- 5.2% (N = 24) and 40.4 +/- 8.8% (N = 16), respectively (P < 0.0001, Student's t-test). In conclusion, the vibratory microinjection method is a new, useful option for gene transfer because it resulted in much less cellular deformation, therefore implicating less cellular damage.

Lupeol esters from the twig bark of Japanese pear (Pyrus serotina Rehd.) cv. Shinko.

Five new lupeol esters, lup-20(29)-ene-3beta-yl eicosanoate, docosanoate, tetracosanoate, hexacosanoate and octacosanoate, were isolated as a mixture from the twig bark of Japanese pear (Pyrus serotina Rehd.) cv. Shinko, together with lupeol and epifriedelinol. Their structures were determined by spectral analyses including 2D-NMR experiments.

Muscle contraction and relaxation described by tactile stiffness.

We developed a tactile sensor system that measures the stiffness of objects (tactile stiffness) and used it to describe the time course of muscle contraction and relaxation. We examined fatigue resistance of the latissimus dorsi muscle (LDM), which is preconditioned for cardiomyoplasty. Time to peak, ripple of LDM, and time constant were calculated from the time course of LDM contraction and relaxation as described by tactile stiffness. We compared conditioned and unconditioned LDMs using these 3 parameters. The time course can be described by tactile stiffness. Tactile stiffness fell exponentially during LDM relaxation. In mean values, time to peak increased 230%, ripple decreased 20%, and time constants increased 424%. Significant differences were shown in 3 parameters between conditioned and unconditioned LDMs (p < 0.05). Our tactile sensor system can describe the time course of LDM contraction and relaxation. Examining the difference in time courses, we might detect the level of LDM preconditioning for cardiomyoplasty.

CD28 as a molecular amplifier extending TCR ligation and signaling capabilities.

Evidence has gathered that CD28 costimulation facilitates T cell activation by potentiating TCR intrinsic-signaling. However, the underlying molecular mechanism is largely unknown. Here we show that, by enhancing T cell/APC close contacts, CD28 facilitates TCR signal transduction. Moreover, the signal supplied by CD28 does not lead to increased Zap-70 and Lat phosphorylation, but amplifies PLCgamma1 activation and Ca(2+) response. We provide evidence that the PTK Itk controls the latter function. Our data suggest that CD28 binding to B7 contributes to setting the level of TCR-induced phosphorylated Lat for recruiting signaling complexes, whereas the CD28 signal boosts multiple pathways by facilitating PLCgamma1 activation. These results should provide a conceptual framework for understanding quantitative and qualitative aspects of CD28-mediated costimulation.

Use of tactile stiffness to detect fatigue in the latissimus dorsi muscle.

In clinical settings, no method has been established to examine the fatigue of a latissimus dorsi muscle (LDM) preconditioned for cardiomyoplasty. We examined the feasibility of measuring muscle stiffness (tactile stiffness) to evaluate muscle fatigue in situ using our tactile sensor. We stimulated canine LDM with burst pacing and monitored both stiffness and tension to determine their relationship. In both dissected LDM and LDM in situ, the decrements of these parameters during burst pacing were compared between preconditioned and unconditioned LDM. In measurement in situ, the sensor probe was placed on the LDM through a small incision. Strong statistical correlation was shown between stiffness and tension (r = 0.935). In decrements of stiffness in situ, there were statistically significant differences between preconditioned and unconditioned LDM. Our tactile sensor system can provide an efficient method for evaluating fatigue of muscles in situ without measuring muscle tension.

Objective evaluation of liver consistency to estimate hepatic fibrosis and functional reserve for hepatectomy.

BACKGROUND: The empiric evaluation of liver consistency is currently used to plan the surgical strategy. The aim of this study was to verify the feasibility of the objective measurement of liver consistency and to check its correlation with liver fibrosis and liver functional reserve. STUDY DESIGN: Fifty-two consecutive patients who underwent hepatic resections in our department were enrolled. The indications for liver resection were hepatocellular carcinoma in 36 patients, metastatic liver tumors in 12 patients, and other conditions in 4 patients. Liver consistency was measured with a new tactile sensor. A fibrosis index was calculated as an expression of the percentage of fibrotic tissue. Liver consistency was compared with the degree of liver fibrosis observed in histologic specimens (fibrosis index) and with liver function parameters. RESULTS: Liver stiffness showed a significant positive correlation with fibrosis index (r = 0.887, p < 0.0001). Liver stiffness also showed significant positive correlation with the indocyanine green test (r = 0.631, p < 0.0001) by a univariate analysis. The indocyanine green test and platelet count were independently and significantly associated with liver stiffness by a multiple regression analysis. In five patients, the liver stiffness values measured intraoperatively differed markedly from those expected from the indocyanine green test values. In these patients, the operative procedures were finally selected based on the liver stiffness measured with the tactile sensor and good clinical outcomes were obtained. CONCLUSIONS: These results show for the first time that liver stiffness can be clinically assessed quantitatively by means of the tactile sensor. The tactile sensor adequately estimates liver stiffness and this estimation is well correlated with liver fibrosis and functional reserve. Liver consistency determined objectively in this manner may be useful for optimizing surgical decision making.

Myocardial tactile stiffness during acute reduction of coronary blood flow.

BACKGROUND: Evaluation of regional myocardial contractile function is of clinical importance. We have developed a new tactile sensor system for accurate measurement of myocardial stiffness in situ. We found that the myocardial stiffness measured by this sensor, which we call tactile stiffness, can be a very useful index for accurate quantification of regional myocardial function. In this study, we used a coronary stenosis model to investigate regional myocardial tactile stiffness under conditions of reduced coronary blood flow. METHODS: The myocardial tactile stiffness, coronary blood flow, and ventricular pressure and volume, of five open chest mongrel dogs weighing 15 to 17 kg, were measured. After measuring the baseline myocardial stiffness, coronary arterial stenosis was induced with a balloon occluder. RESULTS: Reducing the coronary flow to 50% and 25% of the baseline level reduced the end-systolic tactile stiffness significantly from 2.20+/-0.16 g/mm2 to 2.05+/-0.20 g/mm2 (p<0.05) and from 2.21+/-0.16 g/mm2 to 1.96+/-0.18 g/mm2 (p<0.01), respectively. Reducing the flow, to 50% and 25%, increased the end-diastolic stiffness significantly from 1.29+/-0.15 g/mm2 to 1.39+/-0.14 g/mm2 (p<0.01) and from 1.30+/-0.16 g/mm2 to 1.46+/-0.14 g/mm2 (p<0.05), respectively. CONCLUSIONS: We consider that the regional myocardial tactile stiffness will be a useful index sensitive enough to detect changes in regional contractile function under conditions of reduced coronary blood flow.

Molecular cloning and splicing isoforms of mouse p144, a homologue of CA150.

We previously characterized p144 bearing N-acetylglucosamine residues in a rat liver nuclear matrix fraction. Based on partial amino acid sequences of rat p144, mouse p144 cDNA was cloned and sequenced, and its amino acid sequence was predicted. The sequence revealed that p144 is a rat homologue of CA150, which is a transcription factor involved in Tat-activated human immunodeficiency virus type 1 transcription. The reported human CA150 consists of 1098 amino acids and has a leucine zipper-like motif in its carboxyl-region. However, a clone of mouse p144 cDNA encoded a CA150 consisting of 1,034 amino acids. The mouse CA150 was shorter by 64 amino acids than hitherto known human CA150 and lacked the leucine zipper-like motif. We designated the longer and shorter CA150 species as CA150a and CA150b, respectively. The partial nucleotide sequences of other mouse p144 cDNA clones were examined and it was found that some clones encode CA150a having a leucine zipper-like motif. It was suggested that CA150a and CA150b are splicing isoforms. All rat and mouse tissues examined contained transcripts for both CA150a and CA150b. Both transcripts were detected in human blood and Jurkat cells as well as mouse CD4(+) T-cells, which are the HIV-1-sensitive counterpart in humans.

Interaction between A beta(1-42) and A beta(1-40) in Alzheimer's beta-amyloid fibril formation in vitro.

We analyzed the interaction of two kinds of amyloid beta-peptides (A beta), i.e., A beta(1-42) and A beta(1-40), in the kinetics of beta-amyloid fibril (fA beta) formation in vitro, based on a nucleation-dependent polymerization model using fluorescence spectroscopy with thioflavin T. When 25 microM A beta(1-42) was incubated with increasing concentrations of amyloidogenic A beta(1-40), the time to proceed to equilibrium was extended dose-dependently. A similar inhibitory effect was observed when 45 microM A beta(1-40) was incubated with increasing concentrations of A beta(1-42). On the other hand, when 50 microM of nonamyloidogenic A beta(1-40) was incubated with A beta(1-42) at a molar ratio of 10:1 or 5:1, A beta(1-42) initiated fA beta formation from A beta(1-40). The lag time of the reaction shortened in a concentration-dependent manner, with A beta(1-42). We next examined the seeding effect of fA beta formed from A beta(1-42) (fA beta(1-42)) on nonamyloidogenic A beta(1-40). When 50 microM of nonamyloidogenic A beta(1-40) was incubated with 10 or 20 microg/mL (2.2 or 4.4 microM) of fA beta(1-42), the fluorescence showed a sigmoidal increase. The lag time of the reaction was shortened by fA beta(1-42) in a concentration-dependent manner. However, the time to proceed to equilibrium was much longer than when an equal concentration of fA beta formed from A beta(1-40) (fA beta(1-40)) was added to A beta(1-40). The fluorescence increased hyperbolically without a lag phase when 25 microM A beta(1-42) was incubated with 10 or 20 microg/mL (2.3 or 4.6 microM) of fA beta(1-40), and proceeded to equilibrium more rapidly than without fA beta(1-40). An electron microscopic study indicated that the morphology of fA beta formed is governed by the major component of fresh A beta peptides in the reaction mixture, not by the morphology of preexisting fibrils. These results may indicate the central role of A beta(1-42) for fA beta deposition in vivo, among the different coexisting A beta species.

Stimulus-coupled interaction of tyrosine hydroxylase with 14-3-3 proteins.

Tyrosine hydroxylase (TH) is phosphorylated by CaM kinase II and is activated in situ in response to a variety of stimuli that increase intracellular Ca(2+). We report here, using baculovirus-expressed TH, that the 14-3-3 protein binds and activates the expressed TH when the enzyme is phosphorylated at Ser-19, a site of CaM kinase II-dependent phosphorylation located in the regulatory domain of TH. Site-directed mutagenesis showed that a TH mutant in which Ser-19 was substituted by Ala retained enzymatic activity at the same level as the non-mutated enzyme, but was a poor substrate for CaM kinase II and did not bind the 14-3-3 protein. Likewise, a synthetic phosphopeptide (FRRAVpSELDA) corresponding to the part of the TH sequence, including phosphoSer-19, inhibited the interaction between the expressed TH and 14-3-3, while the phosphopeptide (GRRQpSLIED) corresponding to the site of cAMP-dependent phosphorylation (Ser-40) had little effect on complex formation. The complex was very stable with a dissociation constant of 3 nM. Furthermore, analysis of PC12nnr5 cells transfected with myc-tagged 14-3-3 showed that 14-3-3 formed a complex with endogenous TH when the cultured cells were exposed to a high K(+) concentration that increases intracellular Ca(2+) and phosphorylation of Ser-19 in TH. These findings suggest that the 14-3-3 protein participates in the stimulus-coupled regulation of catecholamine synthesis that occurs in response to depolarization-evoked, Ca(2+)-dependent phosphorylation of TH.

In vitro nuclear assembly with affinity-purified nuclear envelope precursor vesicle fractions, PV1 and PV2.

Nuclear envelope precursor vesicles were affinity purified from a Xenopus egg extract by a chromatin binding method. Vesicles bound to chromatin at 4 degrees C were dissociated with a high salt buffer and further fractionated into nuclear envelope precursor vesicle fractions 1 (PV1) and 2 (PV2) by differential centrifugation. PV1 contained larger vesicles. When chromatin was incubated in a Xenopus egg cytosol fraction supplemented with PV1, vesicles bound to chromatin, fused with each other, formed a bilayered nuclear envelope, and assembled into spherical small nuclei. However, the thus assembled nuclei did not grow to the normal size. Nuclear pore complexes were not found on the thus assembled nuclei. On the other hand, PV2 contained smaller vesicles. PV2 vesicles bound to chromatin, fused little with each other in the Xenopus egg cytosol fraction, and no nuclei were assembled. When PV1 supplemented with PV2 was used for the nuclear assembly reaction, the assembled nuclei grew to the normal size. Nuclear pore complexes existed in the thus assembled nuclear envelopes. These results suggested that 1) two vesicle populations, PV1 and PV2, are necessary for the assembly of normal sized nuclei, 2) PV1 contains a chromatin targeting molecule(s) and membrane fusion machinery, 3) PV2 contains a chromatin targeting molecule(s) and a molecule(s) necessary for nuclear pore complex assembly, and 4) PV1 has the ability to assemble a nuclear membrane, and PV2 is necessary for the assembly of nuclear pore complexes and for nuclei to grow to the normal size. An in vitro nuclear assembly system constituted with affinity-purified vesicle fractions, PV1 and PV2, was established.

Molecular cloning of mouse p47, a second group mammalian RuvB DNA helicase-like protein: homology with those from human and Saccharomyces cerevisiae.

A 47k protein (p47) in a high-salt buffer extract of a rat liver nuclear matrix fraction was purified by means of a wheat germ agglutinin affinity column, reversed phase HPLC, and SDS-PAGE, and partial amino acid sequences were analyzed. Based on these sequences, the mouse cDNA of the protein was cloned and sequenced, and its amino acid sequence was deduced. Mouse p47 consists of 463 amino acid residues with a molecular weight of 51,112. The amino acid sequences of human and Saccharomyces cerevisiae p47s were also deduced from the nucleotide sequences of "expressed sequence tag" fragments and genomic DNA, respectively. These sequences contain helicase motifs and show homology to bacterial RuvB DNA helicases acting in homologous recombination. They also show homology with the putative mammalian helicases p50/TIP49 and RUVBL1. Comparison of the amino acid sequences of p47 group proteins and those of p50/TIP49 group proteins revealed the p47 group proteins to comprise a group distinct from the p50/TIP49 proteins. Ultracentrifugation and gel filtration analyses showed that p47 in the rat liver cytosol fraction exists as large complexes of 697k.