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Daptomycin (DAP) is widely used in the treatment of methicillin-resistant Staphylococcus aureus (MRSA) infection. The emergence of DAP non-susceptible MRSA strains during therapy is a major concern in clinical settings. Recent studies revealed that MRSA spontaneously reverts to a subsequent methicillin-susceptible S. aureus (MSSA) strain. However, it is not clear whether DAP non-susceptible MRSA has the ability to revert to a susceptible strain. We obtained an MRSA strain pair, DAP non-susceptible strain and subsequent DAP susceptible strain, from a patient. To understand the underlying mechanism by which DAP non-susceptible MRSA reverts to a susceptible strain, we performed genetic and phenotypic analysis in the strain pair. Although whole-genome analysis revealed four missense mutations, including L826F in mprF, in both strains, the net cell-surface charge was similar between the DAP non-susceptible and susceptible strains. However, the thickness of the cell wall was higher in the DAP non-susceptible strain, which was decreased to the same level as the control after reversion to the DAP susceptible strain. Moreover, the non-susceptible strain showed higher mRNA expression of the two-component system (TCS), such as VraSR, yycG and GraS, with the up-regulated transcription levels of cell-wall biosynthesis-related genes. The expression levels of those genes were decreased after reversion to the susceptible strain. These results indicated that DAP non-susceptibility due to up-regulation of the TCS and cell-wall biosynthesis-related genes may be reversible by the discontinuation of DAP, leading to reversion to the DAP susceptible phenotype.

Background Recently, some studies have highlighted proprotein convertase subtilisin-kexin type 9 (PCSK9) inhibitors produce incremental low-density lipoprotein cholesterol (LDL-C) lowering effect. However, it is unknown whether the lipid composition of plaque changes is associated with serum LDL-C reduction due to PCSK9 inhibitors administration. Purpose The purpose of this study was to determine the effects of PCSK9 inhibitor (PCSK9i) on coronary plaque component in patients with a history of coronary artery disease (CAD) assessed by near-infrared spectroscopy intravascular ultrasound (NIRS-IVUS). Methods A total of 67 non-culprit coronary segments were identified in 34 patients. These lesions were analyzed utilizing NIRS-IVUS at baseline and follow-up coronary angiography (CAG). The subjects were divided into two groups according to lipid-lowering treatment; administration of PCSK9i group (PCSK9i: 19 segments, 9 patients) and traditional statin treatment group (Control: 48 segments, 25 patients). The change of lipid-rich plaque distribution between baseline and follow-up NIRS-IVUS was defined as the change of maximal lipid core burden index (LCBI) score for each of the 4-mm longitudinal segments (maxLCBI4mm). Results Mean duration from baseline to follow-up CAG was 239.4±52.4 days in the PCSK9i group and 341.0±84.1 days in the Control group (p Figure 1 Conclusion Compared with traditional statin therapy, PCSK9i treatment resulted in a greater decrease in lipid component in non-culprit coronary plaques. Therefore, PCSK9i may be useful option in preventing from adverse coronary events for the patients with CAD.

A 43-year-old woman with severe aplastic anemia (SAA) received anti-thymocyte globulin and cyclosporin A (CyA) and achieved hematological remission. Although she had maintained hematological remission, the disease relapsed 10 months after arbitrary discontinuance of maintenance therapy with CyA. Resumption of CyA therapy was not effective, and her condition became complicated with progressive sinusitis with bone destruction, which was refractory to antibiotics, antifungal agents, granulocyte colony-stimulating factor, and surgical drainage. Because of the necessity for early neutrophil recovery (to resolve the infection), we proceeded with a combination therapy using allogeneic peripheral blood stem cell transplantation (PBSCT) promptly followed by granulocyte transfusion (GTX) from the same human leukocyte antigen-identical donor rather than carrying out a second immunosuppressive therapy. The patient showed temporal resolution of infection on the second day after a single GTX. Although the patient had pneumonia on day 11, it was resolved promptly after engraftment on day 16. This report suggests the clinical utility of a salvage therapy with allogeneic PBSCT followed by GTX in a particular case of recurrent SAA with refractory infections.

Dry deposition and dew components on pine foliage were measured from 1998 to 2000 on the urban- and mountain-facing sides of Mt. Gokurakuji in order to estimate the effect of anthropogenic activities to dry deposition and dew concentration on the surfaces of pine foliage. A leaf wash experiment was employed to determine the dry deposition rates on the pine foliage. The NO3− and SO42− dry deposition rates per unit surface area of pine foliage were 1.47 and 0.28 μmol m−2 h−1 respectively on the urban-facing side and 0.32 and 0.09 μmol m−2 h−1 on the mountain-facing side. Dry deposition fluxes of N (NO3−+NH4+) and S (SO42−) to the forest floors were 8.4 kg N ha−1 yr−1 and 2.8 kg S ha−1 yr−1 on the urban-facing, and 3.3 kg N ha−1 yr−1 and 1.8 kg S ha−1 yr−1 on the mountain-facing side, respectively. The higher dry deposition fluxes of N and S on the urban-facing side could be attributed to its proximity to traffic roads and the urban area. The concentrations of most ions in the dew were higher on the urban-facing side (U130) than on the mountain-facing side (M430). NO3− and SO42− concentrations in dew at U130 were 802 and 428 μeq l−1, respectively, while at M430 they were 199 and 222 μeq l−1, respectively, suggesting that higher dry deposition rates on the urban-facing side enhanced their concentrations in the dew on this side. The role of dry deposits and subsequently dissolved ones in dew on the needle surfaces is discussed in terms of pine tree damage by atmospheric depositions.

We cloned a DNA fragment from Saccharomyces cerevisiae that complemented the deficiency in high-affinity glutathione transport activity conferred by a gsh11 mutation, and found that the ORF responsible was YJL212c, which had already been designated as OPT1 and HGT1 by others. Northern analysis clearly demonstrated that this ORF, now referred to as OPT1/ HGT1/ GSH11, was induced by sulfur starvation and repressed by adding cysteine to the growth medium. Reporter gene assays showed that a segment spanning the region between positions -371 and -355 was essential for the regulation of this gene. A sequence of 9 nt, CCGCCACAC (from -364 to -356), in this region was shown to be required for protein binding, using an electrophoretic mobility shift assay. Based on these results, we propose that CCGCCACAC comprises the core of a cis-acting element involved in cysteine-responsive gene regulation in S. cerevisiae.

Carbon-coated material was prepared by impregnation of a polymer and successive thermal treatment. Polymers such as phenol resin, polyacrylonitrile, polyvinylchloride and polivinylidene dichloride were used for impregnation and inorganic oxides such as SiO2, Al2O3, ZrO2 and TiO2 were used as supports. The amount of carbon coating was varied arbitrarily by changing the amount of polymer dissolved in the solvent for impregnation. Carbon-coated materials were characterized by TG/DTA, N2 adsorption, mercury porosimetry and elemental analysis. Properties and structure of carbon-coated materials were discussed.

Reciprocal effects among local breeds and additive and heterosis effects were estimated for total milk production per lactation (TLP), lactation length (LL) and daily milk yield (DMY). Procedures for an optimum crossbreeding system were examined by comparing various crossbred combinations. Three data sets were chosen from the total records collected from 1962 to 1997. Data set I with a total of 5,938 records composed of 9 breed groups for reciprocal crosses, data set II with a total of 6,064 records composed of 5 breed groups for additive and heterosis effects and data set III with a total of 10,319 records composed of 42 breed groups for optimum crossbreeding system were analyzed. The least squares ANOVA was used. The reciprocal crosses between Red Sindhi and the Bangladeshi Local showed significant difference. When the Red Sindhi was used as cow, the cross was superior to the opposite in terms of the three performance traits. The results showed that additive breed effects on both TLP and DMY were positive and highly significant between Bos indicus and Bos taurus, whereas the individual heterosis effects were not significant. In general, any local breeds were improved two to three times by crossing with the Friesian once or twice. However, the effect was reduced when the grading up with the Friesian was repeated more than twice. On the other hand, when the local breed was crossed with the other breeds such as Holstein and Jersey, the improvement of TLP was less. Crossing the F1 cows with the Friesian bull gave further improvement. Almost the same trends were found also for LL and DMY. The local breeds and their crosses are raised mainly for draft animals, then it is recommended that the crossbreds between the local cows and Friesian are used for dairy purposes and more than 75% Friesian bloo di s used for beef. (Asian-Aust. J. Anim. Sci. 2002. Vol 15, No. 6 : 777-782)

Background: The aim of this prospective study is to evaluate feasibility of the periareolar injection of contrast agent (Sonazoid (SNZ)) followed by ultrasonography (US) for identification of sentinel lymph node (SLN) in breast cancer patients with clinically negative node. Patients and Methods: Patients with T1-2N0M0 breast cancer were recruited in this study. They received the periareolar injection of SNZ followed by US to identify contrast-enhanced SLN. Fine needle aspiration biopsy (FNAB) was done for each CE-SLN. Then, they underwent SLN biopsy with the conventional method, blue dye and/or radiotracer (B/R). Results: In almost all cases, contrast-enhanced lymphatic vessels were clearly visualized US soon after the periareolar injection of SNZ, and SLN, into which lymphatic flow was draining, was easily identified. The identification rate of SLN was 98% (98/100) by SNZ and 100% (100/100) by B/R. The number of SLNs identified by SNZ (mean per patient, 1.52) was significantly (P < 0.001) lower than that of those by B/R (2.22). Twenty-five patients had at least one metastasis in the SLNs identified by SNZ and/or B/R. In these patients, SLNs (n=39) identified by both SNZ and B/R showed a significantly (P < 0.0001) higher positivity (74.4%) for metastases than those (n=19) identified by B/R alone (21.1%). Conclusion: Identification of SLN by periareolar injection of SNZ followed by US is a technically easy method and the identification rate of SLN was as high as 98%, being comparable to the conventional B/R. SLNs detected by SNZ seem to represent the true SLNs which first receive lymphatic flow from the tumor among the SLNs detected by B/R. Citation Format: Shimazu K, Ito T, Uji K, Miyake T, Motomura K, Noguchi S. Sentinel lymph node biopsy by contrast-enhanced ultrasonography with sonazoid in patients with breast cancer - Prospective multicenter study [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P2-01-27.

The nuclear magnetic dipole moment mu and electric quadrupole moment Q of the beta-emitting 16N(Ipi = 2(-), T(1/2) = 7.13 s) nucleus have been determined for the first time by detecting its beta-NMR in a MgO crystal and beta-NQR (nuclear quadrupole resonance) in a TiO (2) crystal to be /mu/ = (1.9859+/-0.0011) mu(N) and /Q/ = (17.9+/-1.7) mb, respectively. Although the prediction of mu given by the Hartree-Fock calculation agrees well with the experiment, an abnormally small effective charge for neutrons is required to account for the experimental Q.

To determine the usefulness of a ceramic lunate replacement for the wrist joint as a possible treatment for Kienböck's disease, stress distribution of five wrist joint models (a normal wrist, one with the lunate bone removed, one with a coiled tendon replacement, one with a coiled tendon replacement with a ceramic core, and one with a ceramic lunate replacement coated with sheets of tendon) were analysed by the finite element method. The results of finite element method analyses were compared with those of pressure distribution analyses on the wrist joint using pressure-sensitive film and measurement of pressure on the radio-lunate joint with a tactile sensor. Both the coiled tendon model and ceramic core model were unable to resist applied force. Stress and pressure were well-distributed in the wrist joint of the ceramic lunate implant model. These results indicate that the ceramic lunate implant could be expected to prevent carpal collapse.

The nuclear magnetic-dipole moment μ and electric-quadrupole moment Q of the short-lived β-emitting nucleus 19 O (I π =5/2 + ,T 1/2 =27 s ) have been determined by detecting its β-NMR in CaO and TiO2 crystals to be |μ|=(1.53195±0.00007) μ N and |Q|=(3.7±0.4) mb, respectively. Slight deviations of both values from the shell model predictions given by Hartree-Fock calculations demand further experimental and theoretical studies.

The electric quadrupole interaction of 19O(Iπ=(5/2)+, T1/2=27.0 s) in TiO2 single crystal was studied in detail by means of the β-NQR to determine the electric quadrupole moments Q of short-lived β-emitting nuclei 19O and 13O(Iπ=(3/2)-, T1/2=8.6 ms). Two implantation sites were found for the implanted O nucleus and the quadrupole coupling constants of 19O at these sites were determined. We observed FT-NMR of the enriched stable isotope 17O in TiO2 and obtained the electric field gradient (EFG) at the oxygen substitutional site. With this knowledge, we have determined Q(13O)=11.0 ± 1.3 mb and Q(19O)=3.7 ± 0.4 mb. The present results are compared with the theoretical values calculated by the shell model code, OXBASH and by the Hartree–Fock calculation with the realistic potential.

Large abdominal wall defects may require a prosthesis for closure. The aim of our study was to identify the best material for abdominoplasty in pediatric patients. One hundred twenty-eight Wistar KY strain male rats (3 weeks old) were used. All animals underwent celiotomy via a midline skin incision. They were divided into seven groups as follows: the animals in groups 1 through 6 underwent full-thickness abdominal wall excision 3 cm in diameter. The animals in group 1 underwent primary closure. In groups 2 through 6 the defect was closed with prosthetic material. In Group 7, a sham operation was performed. Daily weights were measured. The animals were killed after 3 and 9 weeks. Adhesion scores were assigned for each group. Vicryl mesh resulted in the fewest adhesions and had no effect on weight gain in the developing rats.

The quadrupole moment of the β -emitter 12 N( I π =1 + , T 1/2 =11 ms) has been measured to be Q ( 12 N)=+9.8±0.9 mb by use of a modified β -NMR method. The present result is 3 times smaller than the value which was deduced from the photoproduction cross-section of pions on 12 C, while it is twice the shell-model prediction based on the Cohen-Kurath wave functions in the Woods-Saxon potential.

A novel gene, designated neurorep 1 , was isolated by differential hybridization screening from a complementary DNA library constructed from the rat facial nucleus whose nerve had been transected seven days before sampling. In situ hybridization revealed that this gene was up-regulated in the repair stage after axotomy. The deduced protein, Neurorep 1, consists of 293 amino acid residues, and its molecular mass is approximately 34,000. Protein sequence motif search indicates that this protein has an ecto-5′-nucleotidase consensus sequence at the carboxyl terminal region. In vitro studies showed that Neurorep 1 significantly increased the activity of ecto-5′-nucleotidase, which is considered to be involved in regeneration and repair of the central nervous system. Neurorep 1 might play a significant role in the repair process of nerve tissues by its regulation of ecto-5′-nucleotidase activity.

The magnetic moment of the proton drip-line nucleus C-9(I(sup (pi)) = 3/2, T(sub 1/2) = 126 ms) has been measured for the first time, using the beta-NMR detection technique with polarized radioactive beams. The measure value for the magnetic moment is 1mu(C-9)! = 1.3914 +/- 0.0005 (mu)N. The deduced spin expectation value of 1.44 is unusually larger than any other ones of even-odd nuclei.

To try to explain disability in the wrist after malunited Colles’ fractures, an experiment was designed to evaluate pressure distribution through the wrist joint with varying degrees of angulation of the distal radius. The results of computer simulation analysis of stress concentration using finite element methods were compared with those of pressure distribution studies on the radial articular surface of cadaveric radii in various positions using pressure-sensitive film. Stress existed in volar regions of the radio-lunate joint under normal pressure in the neutral position, but shifted to dorsal regions of the joint, and was concentrated when dorsal angulation reached 30°. We believe that concentrated loads result in pain and may lead to early degenerative joint disease. Early additional reduction of a displaced fragment or a post-fusion osteotomy should be considered when dorsal angulation exceeds 30° in malunited distal radial fractures to decrease abnormal loading across the wrist joint.

Eugene Goldwasser spent 25 years working on the purification of erythropoietin. He eventually succeeded by using a 7-step purification procedure on 2550 liters of urine collected from people with aplastic anemia. His final yield was 8 mg. Goldwasser published these results in the Journal of Biological Chemistry Classic paper reprinted here.

To clarify the mechanism of FSH secretion from the pituitary induced by activin A, we studied the interaction between activin A and gonadal steroids in inducing FSH release from primary cultured female rat pituitary cells in serum-free medium. The basal release of FSH was stimulated by activin A, testosterone (T) and progesterone (P), and T and P also facilitated basal FSH release stimulated by activin A. The GnRH-stimulated FSH release was facilitated by activin A, P and 17β-estradiol (E2) , but suppressed by T. The effect of activin A on GnRH-stimulated FSH release was facilitated by P, but not affected by T or E2. These findings suggested that the interaction between activin A and T or P may be involved in the regulation of FSH secretion during the estrus cycle in rats.

Summary In this study, longitudinal changes of the occlusal force distribution ratio were examined in lower distal extension removable partial dentures with cast circumferential clasps. Occlusal force applied to the denture base and forces transmitted to the retainers were measured on several separate occasions from the insertion of new dentures to about 4 months after. Two rates of loading were chosen. One was simulated mastication (fast loading rate) and the other was 10 Ns-1 (slow loading rate). Location of the loading points were first premolar (PI), second premolar (P2) and first molar (P3) of the denture. The occlusal force distribution ratio to the retainers was calculated when a load of 20N was applied to the loading point. The results are summarized as follows: (1) The occlusal force distribution ratio at fast loading rate on P1 and P2 was changed until 1 or 11/2 months after the insertion of the new dentures, and then became constant. This constant value was 30% on P1, 20% on P2 and 10% on P3. (2) Slow loading rate produced a greater ratio than the fast loading rate on P2 and P3 while there were no remarkable differences in the ratio between both loading rates on P1.

Rabbit morulae were exposed to a vitrification solution-modified PBS [PB1] medium containing 40% ethylene glycol + 18% Ficoll + 0.3 M sucrose (EFS) for 2, 5, or 10 min at 20 degrees C and were vitrified in liquid nitrogen. When morulae were rapidly warmed, 96% had an intact zona pellucida. When embryos were cultured after removal of the mucin coat, high proportions of them formed blastocoel (79-100%), but the percentage of embryos developed to fully expanded blastocysts decreased with increased exposure time 87%, 40%, and 17%). The survival rate of morulae vitrified after removal of the mucin coat was lower than that of mucin-intact embryos. To assess the development potential in vivo, 131 embryos were vitrified after 2 min of exposure to EFS solution; all the embryos were recovered and 120 were transferred to recipients without removal of the mucin coat, resulting in 78 (65%) full-term fetuses or young. This simple method, which yields high survival both in vitro and in vivo, will be of practical use for vitrifying rabbit embryos.

Lipoperoxide concentration in erythrocytes from workers occupationally exposed to lead (mean blood lead concentration 57.1 (SD 17.6) micrograms/dl) was significantly higher than that in controls. It was not different in plasma from the two groups. The activity of superoxide dismutase (SOD) and catalase in erythrocytes from workers exposed to lead was significantly lower than that of control subjects. The effect of lead was also seen in the glutathione concentration of erythrocytes from lead exposed workers, which was reduced to 69% of that found in erythrocytes from control workers. The increase in methaemoglobin content of erythrocytes from workers exposed to lead was less than expected and not significantly different from that of controls. A positive correlation between lipoperoxide concentration in erythrocytes and lead concentration in blood and a negative correlation between glutathione concentration in erythrocytes and blood lead concentration were found. Incubation of erythrocytes for 24 hours at 37 degrees C in the presence of lead (100 micrograms/dl) produced no changes in glutathione and lipoperoxide concentrations, although there was inhibition of activity of SOD (14.3%), catalase (10.1%), and glutathione peroxidase (35.1%). A similar experiment with heparinised whole blood showed increased haemolysis with no changes in membrane lipid peroxidation of erythrocytes. It is postulated that the lowered concentration of glutathione and decreased activity of SOD, catalase, and glutathione peroxidase in erythrocytes from workers exposed to lead may play a part in the increased membrane lipid peroxidation. Furthermore, the results suggest the possibility that leucocytes, or platelets, or both, may induce haemolysis in the presence of lead.

排便時の肛門部激痛を訴え,肛門部に潰瘍を認めた63歳の男性で,潰瘍部の生検から結核性肛門潰瘍と診断し,約2カ月にわたる抗結核治療によって潰瘍の治癒をみた1例を報告した,結核性肛門潰瘍は稀なものであり,肛門癌と誤られやすいが,本症例のごとく,肺結核患者であること,潰瘍の性状には癌による場合とは異なる点がいくつかあることが診断の手掛かりとなる.結核性肛門潰瘍は抗結核治療によく反応するので,生検によって正確な診断を下し,不必要な手術を避けるべきである.

We report the results of pathologic and biochemical studies in a patient with 6q-linked autosomal recessive juvenile parkinsonism (AR-JP). Neuronal loss and gliosis were restricted to the substantia nigra and the locus ceruleus. No Lewy bodies were found, but neurofibrillary tangles and argyrophilic astrocytes were seen in the cerebral cortex and brainstem nuclei. The later findings, which have not been reported previously in AR-JP, suggest the pathologic heterogeneity of 6q-linked AR-JP.

Overexpression and/or gene amplification of c-Src and members of the epidermal growth factor receptor (EGFR/ErbB) family have been implicated in the pathogenesis of breast cancer. Although members of the EGFR family are known to form heterocomplexes with one another, c-Src has also been shown to physically interact with members of this family in breast cancer cell lines and tumors. This paper investigates the role of c-Src in modulating the physical and functional interaction between ErbB2 and ErbB3, two family members that preferentially associate with one another and together exhibit high oncogenic potential. We show that overexpressed wild-type c-Src enhances heterocomplex formation of ErbB2 and ErbB3 that results in increased basal and/or heregulin-induced activation of receptors, and their downstream intracellular effectors. Expression of a kinase-inactive form of c-Src (K(-) c-Src) or pharmacological inhibition of c-Src by PP2 negatively affects these events. Furthermore, cellular motility and anchorage-independent growth promoted by the ErbB2/ErbB3 heterocomplex are dependent upon c-Src, as demonstrated by the effects of K(-) c-Src overexpression or treatment with PP2. In contrast to previous studies that defined a role for c-Src downstream of ErbB2/ErbB3, the current work suggests an upstream mechanism, whereby c-Src enhances ErbB2/ErbB3 signaling and biological functions by positively modulating the association between ErbB2 and ErbB3.

In this research, a redundant driving system seen in the animate was imitated. The redundant driving system composed of a standard joint of rigid body, a 3-DOF ankle joint and a redundant cable were designed and the effectiveness of the cable control mechanism was confirmed. Moreover, a crutch was incorporated to this mechanism and we studied the complete dynamic walking in which the weight characteristic was also used.

In phase 1 eight standard pregnant Wistar rats were used. After delivery of a litter, each mother underwent splenectomy and autotransplantation of a piece of her spleen, which has been divided into eight pieces of equal size in the omentum. The weaning rats were divided into four groups (A–D). Rats in group A (N 5 18) underwent splenectomy, and each received a transplanted piece of the mother’s spleen. Group B (N 5 18) rats each received a transplanted piece of the mother’s spleen. Group C rats (N 5 21) underwent splenectomy only, and group D rats (N 5 20) underwent sham operations. All animals were anesthetized with pentobarbital (40 mg/kg IP) for surgical intervention. Neither immunosuppressive agents nor antibiotics were administered. The animals were weighed daily and underwent extirpation of the native spleen and transplanted spleen to measure the organ weight and evaluate the organ histologically and immunologically at 4 weeks or 6 months after surgery. In phase 2 another group of rats was used to evaluate the effects of combining splenectomy and splenotransplantation on tolerance to other transplants. Weaning rats were divided into four groups and prepared as in phase 1; in addition, they underwent skin transplantation from the mother to the offspring’s back, and the condition of the transplanted skin was evaluated daily and immunohistologic studies were conducted. Histological evaluation was done using hematoxylin and eosin stain.

Formation and inhibition of malonaldehyde (MA) from blood plasma lipids oxidized by Fenton's reagent in the absence or presence of probucol [4,4′-(isopropylidenedithio)bis(2,6-di- tert -butylphenol)] and l -ascorbic acid were investigated. The amount of MA formed was quantitatively analysed by gas chromatography. l -Ascorbic acid inhibited MA formation by about 30% at the level of 4.0 μ mol, but the amount of MA formed was increased by the presence of probucol. When 3.0 μ mol oxidized probucol was hydrolysed at pH 1, 3 and 5, 2616.5 nmol, 287.5 nmol and 103.9 nmol MA were recovered, respectively. This is the first report of quantitative analysis of MA formed from probucol on oxidation.

We examined the effects of PACAP on growth hormone (GH) secretion, intracellular Ca2+ concentrations ([Ca2+]i) and ion channels in GH3 cells. PACAP-38 and vasoactive intestinal polypeptide (VIP) (10−9M) increased GH secretion. PACAP-induced increase in GH secretion was inhibited by PACAP(6–38), a PACAP antagonist. PACAP-38 (10−10M) increased [Ca2+]; in GH3 cells. The increase in [Ca2+]; was inhibited by 1) PACAP(6–38) (10−6M), 2) nicardipine (10−5M), a Ca2+ channel blocker, 3) depletion of extracellular Ca2+ or 4) tetrodotoxin (3x10−7M), a Na+ channel blocker. Na+ channel currents were increased by PACAP, whereas Ca2+ channel currents were unchanged. 8-bromo-cAMP (10−3M) increased [Ca2+]i and Rp-cAMPS (10−3M), an inhibitor for protein kinase A, eliminated PACAP-induced increase in [Ca2+]i. Thus the present study suggests that PACAP activates Na+ channels via cAMP-protein kinase A system coupled with its receptor to depolarize membrane potential, which in turn activates Ca2+ channels to induce an increase in [Ca2+]i, a trigger for GH secretion.

Background The prognosis of gastric cancer with peritoneal metastasis is extremely poor. Neither systemic chemotherapy nor surgery alone prolongs survival of patients significantly. Methods Patients diagnosed with advanced gastric cancer underwent staging laparoscopy and received chemotherapy when peritoneal dissemination and/or cancer cells on peritoneal cytology were confirmed. The chemotherapy regimen consisted of S-1, weekly intravenous and intraperitoneal paclitaxel, which was verified in our phase II trial (Ann Oncol 2009). S-1 was administered at 80 mg/m2/day for 14 consecutive days, followed by 7 days rest. Paclitaxel was administered intravenously at 50 mg/m2 and intraperitoneally at 20 mg/m2 on days 1 and 8. Clinical response of chemotherapy was assessed by computed tomography, gastroendoscopy, peritoneal cytology and second-look laparoscopy. Radical gastrectomy was carried out when macroscopic curative resection was made achievable by chemotherapy. Chemotherapy was restarted after operation as soon as possible. Overall survival, relapse free survival, morbidity and mortality of gastrectomy were evaluated. Results Out of 100 patients with peritoneal metastasis who received chemotherapy, 60 patients underwent gastrectomy after response to chemotherapy, including 54 with macroscopic metastasis and 6 with positive peritoneal cytology only. A median of three courses were administered preoperatively (range 1–16). Total or distal gastrectomy with lymphnode dissection was carried out in 54 or 6 patients, respectively. The median survival time was 34.5 months. The median relapse-free survival was 16.7 months. The first site of relapse was the peritoneum in 24 patients and the other organ site in 17 patients. Postoperative complications included anastomotic leakage and pancreatic fistula in two patients each, which were healed conservatively. There were no treatment-related deaths. Conclusions Gastrectomy combined with S-1, intravenous and intraperitoneal paclitaxel is safe and active for gastric cancer patients with peritoneal metastasis.

e15585 Background: For ovarian cancer patients, it remains unclear whether paclitaxel (T) or docetaxel (D) should be administered intraperitoneally (IP) or intravenously (IV). Because it is virtual...

The purpose of this study was to perform longitudinal measurement of forces transmitted from the denture base to retainers of lower distal-extension removable partial dentures with a conus crown telescopic system. The experimental denture was embedded with a force-detecting unit which could detect a change in lateral and vertical forces transmitted from denture base to retainers at the same time. Forces were measured during chewing of food on several separate occasions from the insertion of new dentures to about 3 months after. Max.ver (maximal mean value of vertical force) and Max.lat (maximal mean value of lateral force) were calculated from the peak level of all strokes of chewing of food at the minimum of the interocclusal distance. The results are summarized as follows: (i) There were no significant changes of Max.ver and Max.lat; (ii) There were no significant differences of Max.ver between subjects the value of which was about 20 N; (iii) There were significant differences of Max.lat between subjects the value of which was less than 15 N.

Elements of the vertebrate skeleton are initiated as cell condensations, collectively termed the 'membranous skeleton' whether cartilages or bones by Gruneberg (1963). Condensations, which were identified as the basic cellular units in a recent model of morphological change in development and evolution (Atchley and Hall 1991) are reviewed in this paper. Condensations are initiated either by increased mitotic activity or by aggregation of cells towards a centre. Prechondrogenic (limb bud) and preosteogenic (scleral ossicle) condensations are discussed and contrasted. Both types of skeletogenic condensations arise following epithelial-mesenchymal interactions; condensations are identified as the first cellular product of such tissue interactions. Molecular characteristics of condensations are discussed, including peanut agglutinin lectin, which is used to visualize prechondrogenic condensations, and hyaluronan, hyaladherins, heparan sulphate proteoglycan, chondroitin sulphate proteoglycan, versican, tenascin, syndecan, N-CAM, alkaline phosphatase, retinoic acid and homeo-box-containing genes. The importance for the initiation of chondrogenesis or osteogenesis of upper and lower limits to condensation size and the numbers of cells in a condensation are discussed, as illustrated by in vitro studies and by mutant embryos, including Talpid3 in the chick and Brachypod, Congenital hydrocephalus and Phocomelia in the mouse. Evidence that genes specific to the skeletal type are selectively activated at condensation is discussed, as is a recent model involving TGF-beta and fibronectin in condensation formation. Condensations emerge as a pivotal stage in initiation of the vertebrate skeleton in embryonic development and in the modification of skeletal morphology during evolution.

Using the Drosophila cell line MLDmBG-1, a monoclonal antibody aBG-1 that can inhibit not only cell clumping but also cell spreading was generated. This antibody immunoprecipitates a complex of molecules consisting of a major 120 × 10(3) Mr and other components. To characterize the 120 × 10(3) Mr component, we purified it, generated antibodies to it, and cloned its cDNA. Sequencing of this cDNA suggests that the 120 × 10(3) Mr molecule is identical to PS beta, a beta chain of Drosophila integrins. The other components immunoprecipitated included two alpha chains of Drosophila integrins, PS1 alpha and PS2 alpha, as revealed using specific antibodies to these molecules. These suggest that aBG-1 recognizes the PS beta associated with PS1 alpha or PS2 alpha. However, immunostaining of embryos and larvae with aBG-1 showed that the staining pattern is similar to that for PS2 alpha but not for PS beta, suggesting that the antibody preferentially recognizes the PS beta associated with particular alpha chains in situ. We then attempted to characterize the ligands for these integrin complexes, using culture dishes coated with various vertebrate matrix proteins. These cells spread very well on dishes coated with vitronectin and, to a lesser extent, on those with fibronectin. This spreading was partially inhibited by aBG-1, but not by other control antibodies or RGD peptides. The cell attachment to these substrata was not affected by the antibody. The cells also can attach to dishes coated with laminin but without spreading, and this attachment was not inhibited by aBG-1. Furthermore, they do not attach to dishes coated with collagen type I, type IV, and fibrinogen. These results indicate that Drosophila PS integrins can recognize vertebrate vitronectin, and also fibronectin with a weaker affinity, at sites other than RGD sequences, and thus can function in cell-substratum adhesion.

Many derivatives of the nprM promoter of Bacillus stearothermophilus and the strong early promoter, A3, of coliphage T3 were designed and chemically synthesized. These promoters consisted of some or all of the AT box, consensus sequence, tac promoter sequence, spacer, and lac operator. The promoter activities were assessed by their ability to express the cat gene in Escherichia coli. One of the derivatives of the A3 promoter, which contained the lac operator, was much stronger (about 3.5 times) than the tac promoter. The promoter activities in E. coli were considerably modified by the substitutions in the -43 (AT box) and -35 regions.

The effects of ascorbic acid (AsA)-deficiency on the development of mammary glands were investigated using mutant rats (osteogenic disorder syndrome rats; ODS rats) with hereditary inability to synthesize AsA. Female ODS rats of 21 days old were castrated and divided into two groups. One group was given AsA in their drinking water, and the other was not. All the rats received a daily injection of oestradiol-17 beta and progesterone (EP) from day 28 to day 49 of age. After EP treatment, the concentrations of AsA in the mammary glands of rats not given AsA were less than one tenth of those of rats given AsA and the contents of hydroxyproline in the mammary glands of the former rats were about half of those in the latter. Furthermore, the concentration of serum prolactin in rats not given AsA was reduced to about one third of that in rats given AsA. After EP treatment, whole mounts of mammary glands showed that in rats not given AsA the development of ducts was impaired and there was extensive accumulation of endbuds. Consistent with this finding, EP injections did not increase the area of parenchyma in the mammary glands of rats not given AsA, whereas they increased it about 2-fold in rats given AsA. Moreover, after EP treatment the amount of alpha-lactalbumin was significantly less in the mammary parenchyma of rats not given AsA than in that of rats given AsA. On the other hand, AsA deficiency did not impair the response of the mammary cells to insulin or prolactin in terms of DNA synthesis and alpha-lactalbumin production. These findings indicate that AsA deficiency impaired the development of mammary glands. This effect may be partly attributable to a defect in collagen synthesis in the mammary glands and a decrease in the concentration of serum prolactin.

13162 Background: Focal adhesion kinase (FAK) is a non-receptor cytoplasmic tyrosine kinase that regulates multiple cell functions. Elevated expression levels of FAK have been detected in various tumor samples and are closely correlated with invasive potential. Activation of integrins and the growth factor receptors result in FAK autophosphorylation at Y397 and the presentation of suitable binding sites for proteins containing either SH2 or phosphotyrosine binding domains. Recent evidences suggest that FAK plays important roles in cancer cell proliferation and survival. IGF-IR function is required for tumor cell survival, but dispensable for survival of normal cells. Therefore, a dual inhibitor of both kinases may selectively block the growth, migration, and survival of FAK- and IGF-IR- expressing tumor cells compared to proliferating and migrating normal cells. Methods: In this study, anti-cancer activity of NVP-TAE226 that is identified as a potent and selective FAK inhibitor was evaluated in cancer cell lines panel and MIA PaCa-2 pancreatic carcinoma in vivo model. Results: Mean GI50 value of NVP-TAE226 against 37 cancer cell lines was 0.76 μmole/L. Inhibition of cancer cell proliferation was not affected by expression of P-glycoprotein, suggesting that NVP-TAE226 is not served as a substrate of P-glycoprotein. Oral administration of NVP-TAE226 efficiently inhibited MIA PaCa-2 human pancreatic tumor growth at all doses tested. Tumor stasis was observed at a dose of 30 mg/kg, qd for 7×/week and tumor regression was observed at a dose of 100 mg/kg, qd for 5×/week. All animals tolerated NVP-TAE226 treatment up to 100 mg/kg, 5×/wk, qd, po for 2 weeks with no body weight loss. Inhibition of downstream signaling such as phosphorylation of Akt at Serine473 was accompanied by inhibition of FAK phosphorylation in human pancreatic carcinoma cell lines. Conclusions: NVP-TAE226 is a novel class of selective and small molecule kinase inhibitors with a potent in vivo activity and potential therapeutic application. No significant financial relationships to disclose.