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This paper describes our approach that is built upon the potential energy surface (PES)-based conformational analysis. This approach automatically deduces a conformational transition network, called a conformational reaction route map (r-map), by using the Scaled Hypersphere Search of the Anharmonic Downward Distortion Following method (SHS-ADDF). The PES-based conformational search has been achieved by using large ADDF, which makes it possible to trace only low transition state (TS) barriers while restraining bond lengths and structures with high free energy. It automatically performs sampling the minima and TS structures by simply taking into account the mathematical feature of PES without requiring any a priori specification of variable internal coordinates. An obtained r-map is composed of equilibrium (EQ) conformers connected by reaction routes via TS conformers, where all of the reaction routes are already confirmed during the process of the deduction using the intrinsic reaction coordinate (IRC) method. The postcalculation analysis of the deduced r-map is interactively carried out using the RMapViewer software we have developed. This paper presents computational details of the PES-based conformational analysis and its application to d-glucose. The calculations have been performed for an isolated glucose molecule in the gas phase at the RHF/6-31G level. The obtained conformational r-map for α-d-glucose is composed of 201 EQ and 435 TS conformers and that for β-d-glucose is composed of 202 EQ and 371 TS conformers. For the postcalculation analysis of the conformational r-maps by using the RMapViewer software program we have found multiple minimum energy paths (MEPs) between global minima of

Epidemiological and pathological studies of Helicobacter spp. in canine stomachs in Japan were performed to investigate strain specific pathogenicity. Gastric biopsies from 144 dogs with gastrointestinal diseases were evaluated for the presence of Helicobacter spp. using genus and species specific PCRs for Helicobacter felis, Helicobacter bizzozeronii, Helicobacter heilmannii sensu stricto (s.s.) and Helicobacter pylori. PCR indicated that 50/144 (34.7%) dogs were infected with Helicobacter spp. Of the genus positive samples, 21/50 could not be amplified by any of the species specific PCRs. To investigate Helicobacter at the species level, partial ureAB gene sequences from 48/50 genus positive samples were determined; 47 strains were identified. Thirty-five strains from 45 cases were closely related to H. heilmannii s.s. (89–99% sequence similarity), seven strains from seven cases were closely related to H. bizzozeronii (95–99% sequence similarity), three strains from three cases were closely related to Helicobacter felis (86%, 98% and 99% sequence similarity), one strain from one case was closely related to Helicobacter salomonis (99% sequence similarity) and one strain from one case was closely related to H. pylori (99% sequence similarity). Dogs infected with Helicobacter spp. most similar to H. heilmannii s.s. had a higher frequency of moderate to severe gastritis than dogs negative for Helicobacter spp. (P = 0.044). In conclusion, the predominant Helicobacter spp. detected in canine stomachs in our study were most closely related to H. heilmannii s.s. and displayed substantial genetic diversity. Infection with Helicobacter spp. may be associated with more severe gastritis in dogs.

Serum amyloid A (SAA) is one of the major acute phase proteins in cats and humans. SAA concentrations increase in response to the inflammatory status and secondary amyloid A amyloidosis has been documented in cats. In order to control the SAA concentration, it is important to clarify how the SAA protein is metabolized. Although the details of SAA metabolism in the body remain unknown, human and murine research indicates that macrophages play a key role in SAA uptake. The objectives of this study were to demonstrate SAA uptake by feline macrophages and to evaluate the effects of lipopolysaccharide (LPS) and dexamethasone (Dex) on SAA uptake. The concentration of recombinant feline SAA added to a feline macrophage culture was decreased in a time-dependent manner and was significantly reduced after a 24-h incubation, as demonstrated by enzyme linked immunosorbent assay (ELISA). SAA uptake into feline peripheral macrophages was demonstrated by immunofluorescence microscopy. Pretreatment to macrophages with LPS did not affect this decrease in the SAA concentration, but this was significantly blocked by Dex pretreatment. In conclusion, SAA was incorporated by feline macrophages and pretreatment with Dex inhibited SAA uptake by macrophages in this study. Further investigation is needed to determine the molecules that influence SAA uptake by macrophages and the effect of clinical glucocorticoid usage on the SAA concentration in cats.

In theoretical studies of chemical reactions involving multiple potential energy surfaces (PESs) such as photochemical reactions, seams of intersection among the PESs often complicate the analysis. In this paper, we review our recipe for exploring multiple PESs by using an automated reaction path search method which has previously been applied to single PESs. Although any such methods for single PESs can be employed in the recipe, the global reaction route mapping (GRRM) method was employed in this study. By combining GRRM with the proposed recipe, all critical regions, that is, transition states, conical intersections, intersection seams, and local minima, associated with multiple PESs, can be explored automatically. As illustrative examples, applications to photochemistry of formaldehyde and acetone are described. In these examples as well as in recent applications to other systems, the present approach led to discovery of many unexpected nonadiabatic pathways, by which some complicated experimental data have been explained very clearly.

We used life cycle assessment to compare alternative technologies for the disposal of sewage sludge and food waste with the aim of identifying the preferred methods for reducing greenhouse gas (GHG) emissions, recovering phosphorus (P), and minimizing risks to public health. We examined two systems: (i) traditional incineration of food waste and separate treatment of sewage sludge, and (ii) a proposed new system in which domestic food waste is mixed with sewage sludge in an anaerobic digestion tank at a sewage treatment plant. We examined variations to each of these systems as alternative technologies for the processing of sludge: low-temperature incineration, high-temperature incineration, composting, production of cement feedstock, low-temperature carbonization, dry granulation, and pyrolysis gasification. We also evaluated the effect of different P recovery technologies: the calcium hydroxyapatite (HAP) method, the magnesium ammonium phosphate (MAP) method, alkaline extraction, and partial-reduction melting. A functional unit of 100,000 people receiving disposal services was used as the basis for life cycle assessment calculations. We found that for each of the technologies compared, the combined sludge and food waste digestion system was superior to separate treatment and disposal. Among the various technology configurations of the combined system, two were judged to be superior to the others with respect to both GHG emissions and P recovery: (i) the pyrolysis gasification of sludge with the MAP method and alkaline extraction from ash for P recovery (Y-5); and (ii) composting of sludge and the MAP method for P recovery (Y-1). When a detailed consideration of health risks was added to the analysis, configuration Y-5 was identified as the best option to target for technology renewal because of its superior disability adjusted life year (DALY) to that of configuration Y-1, which caused greater heavy metal emissions.

Fractalkine, also known as CX(3)CL1, is a unique chemokine that mediates inflammatory responses and is involved in the pathogenesis of several inflammatory disorders, including inflammatory bowel disease (IBD) in humans. In this study, we isolated cDNAs encoding canine fractalkine and its receptor CX(3)CR1, and assessed the biological activity of these molecules. The deduced amino acid sequence of the canine fractalkine cDNA showed 66% and 57% identity to human and mouse homologs, respectively. The N-terminal chemokine domain of the canine fractalkine showed 68% and 65% identity to human and mouse counterparts, respectively. The canine CX(3)CR1 amino acid sequence showed close homology to its human (83% identity) and mouse (81% identity) counterparts. Fractalkine and CX(3)CR1 mRNA were detected in all tissues in this study. Relatively higher expression levels of fractalkine mRNA were observed in the brain, medulla spinalis, small intestine, and mesenteric lymph nodes (MLNs), whereas higher expression levels of CX(3)CR1 mRNA were observed in the medulla spinalis, brain, liver, small intestine, and MLNs. The cross-reactivities of anti-human fractalkine antibody and anti-rat CX(3)CR1 antibody to canine proteins were confirmed using recombinant canine fractalkine and a cell line overexpressing canine CX(3)CR1, respectively. A transwell chemotaxis assay showed that the recombinant canine fractalkine induced migration in canine lymphoid cells expressing CX(3)CR1. The present study will be useful in understanding the canine immune system and the immunopathogenesis of canine inflammatory diseases.

The chemokine receptor CCR9, which interacts with the thymus-expressed chemokine TECK/CCL25, contributes to the localization of lymphocytes to the small intestine, and is implicated in the development of human inflammatory bowel disease (IBD); however, their role in canine IBD is unknown. The objective of this study was to isolate cDNA encoding CCR9 and to investigate CCR9 expression in normal canine tissues and lymphoid cell lines. The complete open reading frame contained 1104 bp, encoding 367 amino acids, with 85% and 81% identity to human and mouse homologs, respectively. CCR9 mRNA was detected in all tissues investigated with the highest expression level in the small intestine. CCR9 mRNA was also expressed in GL-1, a canine B cell leukemia cell line, but not in CLBL-1, a canine B cell lymphoma cell line. Immunoblot and flow cytometry analyses of these cell lines using an anti-human CCR9 monoclonal antibody revealed that CCR9 protein expression was detected only in GL-1, indicating the cross-reactivity of the antibody. Using the antibody, flow cytometry showed that the proportions of CCR9(+) cells were small (mean, 4.88%; SD, 2.15%) in the normal canine PBMCs. This study will be useful in understanding canine intestinal immunity and the immunopathogenesis of canine IBD.

Biliary sludge in dogs is dismissed commonly as an incidental finding. On the other hand, gallbladder mucocele is reported increasingly in dogs and can lead to biliary obstruction or gallbladder rupture. Cholestasis is suspected to play a role in development of sludge and mucoceles, though there are no data in dogs to support this. We investigated gallbladder emptying, a key factor in biliary flow, in dogs with mobile sludge, immobile sludge, or gallbladder mucocele and in healthy controls. Gallbladder ejection fraction estimated by ultrasonography was used as the index of gallbladder emptying. The ejection fraction at 60 min after eating was significantly decreased in all three abnormal groups. Moreover, all dogs with sludge or a mucocele had gallbladder distension. These changes were the greatest in the mucocele group. Thus, biliary stasis occurs not only in dogs with gallbladder mucocele but also in dogs with biliary sludge. Cholestasis may play a role in the pathogenesis or progression of these diseases in dogs.

We used the bacteriophages Qβ and MS2 to determine whether viruses are inactivated by aluminum coagulants during the coagulation process. We performed batch coagulation and filtration experiments with virus-containing solutions. After filtering the supernatant of the coagulated solution through a membrane with a pore size of 50 nm, we measured the virus concentration by both the plaque forming unit (PFU) and polymerase chain reaction (PCR) methods. The virus concentration determined by the PFU method, which determines the infectious virus concentration, was always lower than that determined by the PCR-based method, which determines total virus concentration, regardless of infectivity. This discrepancy can be explained by the formation of aggregates consisting of several virus particles or by the inactivation of viruses in the coagulation process. The former possibility can be discounted because (i) aggregates of several virus particles would not pass through the 50-nm pores of the filtration membrane, and (ii) our particle size measurements revealed that the virus particles in the membrane filtrate were monodispersed. These observations clearly showed that non-infectious Qβ particles were present in the membrane filtrate after the coagulation process with aluminum coagulants. We subsequently revealed that the viruses lost their infectivity after being mixed with hydrolyzing aluminum species during the coagulation process.

Background Vincristine induces gastrointestinal motility disorders in humans. Adverse gastrointestinal events are commonly observed in dogs receiving vincristine. Objectives To evaluate gastric motility after vincristine administration in dogs and the prophylactic effect of a prokinetic agent, mosapride. Animals Five healthy Beagle dogs. Methods Five dogs received vincristine IV at a dosage of 0.75 mg/m2. The motility index (MI) of the antral contraction was ultrasonographically evaluated 30 minutes postfeeding before administration of vincristine and for 6 days after vincristine treatment. After a 6-week washout period, the dogs received vincristine with mosapride (2 mg/kg PO, q24h for 6 days), and the MI was re-evaluated. Adverse gastrointestinal events were evaluated according to the Veterinary Co-operative Group Common Terminology Criteria for Adverse Events (VCOG-CTCAE). Results After vincristine administration, a significant decrease (P

Lymphoid neoplasms are usually diagnosed on the basis of cytological and histopathological findings. However, in some cases, discrimination of lymphoid neoplasms from reactive lymphoid proliferation is difficult. PCR amplification of complementarity-determining region 3 (CDR3) of the immunoglobulin heavy-chain variable region (IGHV) gene can be used to assess clonality of B-cell populations as a supportive diagnostic tool for B-cell neoplasms. Because of the sequence variation and possible somatic hypermutation of the IGHV gene, sensitivity of the PCR-based assay to detect clonal IGHV gene rearrangement largely depends on the sequences and numbers of primer sets. Prior to the development of an efficient assay, we cloned and sequenced 97 IGHV complementary DNAs (48 IGHV-1 and 49 IGHV-3 clones) from normal cat spleens. On the basis of these sequences, we designed 6 forward primers at the variable region and 5 reverse primers at the joining region. Using each of 6 forward primers and a mixture of 5 reverse primers, we amplified CDR3 of IGHV genes and analyzed the PCR products by conventional PAGE and Genescan analyses using fluorescence-labeled primers. Twenty-six feline B-cell neoplasms diagnosed by histopathological and immunohistochemical examinations were subjected to the newly developed analysis of IGHV gene rearrangement. Clonal IGHV gene rearrangement was detected in 22 of 26 (84%) samples by both PAGE and Genescan analyses. To reduce the number of PCR reactions, we constructed a multiplex PCR analysis system using a mixture of IGHV-1- and IGHV-3-specific primers as forward primers and a mixture of 5 joining region reverse primers. Results of the multiplex PCR were 100% concordant with those obtained by each of the singleplex PCRs. The multiplex PCR-based assay and Genescan analysis developed in the present study would be useful and practical tools to detect clonal IGHV gene rearrangement in feline B-cell neoplasms.

Takeuchi, Y., Fujino, Y., Fukushima, K., Watanabe, M., Nakagawa, T., Ohno, K., Sasaki, N., Sugano, S., Tsujimoto, H. Biological effect of tyrosine kinase inhibitors on three canine mast cell tumor cell lines with various KIT statuses. J. vet. Pharmacol. Therap. 35, 97–104. Tyrosine kinase inhibitors (TKIs) can be important in the treatment of canine mast cell tumor (cMCT). Meanwhile, some TKIs have been identified as substrates for ABCB1. The inhibitory effect of four TKIs (axitinib, imatinib, masitinib, and vatalanib) for proliferation and phosphorylation of c-Kit receptor as well as the expression and function of ABCB1 were investigated in three cMCT cell lines (HRMC, VIMC1, and CMMC1). The IC50 values of the TKIs in HRMC, the only cell line with wild-type KIT, were clearly higher than those in CMMC1 and VIMC1. In HRMC and CMMC1, both the growth and phosphorylation of c-Kit receptor were suppressed proportionally by the TKIs. VIMC1 required higher concentrations for the inhibition of c-Kit receptor phosphorylation than those in cell growth. The treatment with cyclosporine increased the effects of the TKIs on VIMC1 since ABCB1 was expressed in VIMC1. The results indicated that cMCT cell lines harboring wild-type KIT had lower sensitivity to TKIs. The growth of VIMC1 was seemingly reduced by TKIs through the inhibition of other tyrosine kinases than c-Kit receptor. There was little influence of ABCB1 on TKI effects to the proliferation of VIMC1. These results will be helpful to understand the different sensitivity to TKIs in cMCT patients.

We established a novel feline B-cell line, MS4, from the neoplastic pleural effusion of a cat with cutaneous B-cell lymphoma. Immunophenotype staining of the MS4 cells was positive for CD20, CD79α, and IgA and negative for CD3, CD4, CD5, CD8α, CD18, CD21, CD22, IgM, IgG, Ig light chain, and MHC class II. PCR analysis for immunoglobulin heavy chain gene rearrangements revealed a monoclonal rearrangement, whereas no clonal rearrangement of the T-cell receptor γ gene was detected. Southern blotting with an exogenous feline leukemia virus (FeLV) U3 probe revealed no integration of exogenous FeLV provirus. The MS4 cell line is the first FeLV-negative feline B-cell lymphoma cell line, and may be used to investigate the pathogenesis of spontaneously occurring feline lymphoma and the development of new therapies.

BCL6 is known to be a key molecule in germinal center (GC) formation of lymph nodes, and its expression profiles have been implicated in the prognosis of diffuse large B-cell lymphoma in humans. The present study was carried out to characterize canine BCL6 cDNA and to indicate the technical methods for detection of the BCL6 protein in dog tissues. The deduced amino acid sequence of canine BCL6 showed close homology to that of human BCL6 (96.3%), especially in the zinc-finger motifs and POZ (poxvirus and zinc finger) domain with complete identity. Immunoblot analysis of a canine lymph node with an anti-human BCL6 monoclonal antibody revealed a band of 80 kDa. Immunohistochemical staining using the same antibody produced positive reactions in the cells exclusively localized in the GC of a canine lymph node. This study will be useful for the molecular classification of canine B-cell lymphomas with different prognoses.

Background: We developed previously a minimal residual disease (MRD) monitoring system in dogs with lymphoma by exploring a highly sensitive real-time PCR system. Objectives: To identify the change in MRD before clinical relapse in dogs with lymphoma that achieved complete remission after chemotherapy. Animals: Twenty dogs with multicentric high-grade B-cell lymphoma. Methods: MRD levels in peripheral blood mononuclear cells (PBMCs) were measured by real-time PCR amplifying the rearranged immunoglobulin heavy chain gene. MRD measurement and clinical assessment were performed every 2–4 weeks for 28–601 days after completion of chemotherapy. An increase in MRD was defined as an increase by more than 0.5, calculated by log10[copy number of MRD per 105 PBMCs], based on the uncertainty level observed in a canine lymphoma cell line. Results: During the follow-up period, 15 dogs relapsed in 28–320 days (median, 120 days) after completion of chemotherapy. An increase in MRD was detected 2 weeks or more before relapse in 14 of the 15 dogs, but an increase in MRD before relapse could not be detected in the remaining 1 dog. The time from increased MRD to clinical relapse was 0–63 days (median, 42 days). In contrast, no increase in MRD was detected in 5 dogs that did not experience clinical relapse. Conclusion and Clinical Importance: An increase in MRD can be detected before clinical relapse in dogs with lymphoma. Application of early reinduction therapy based on an increase in MRD before clinical relapse may improve treatment outcome in canine lymphoma.

Background: The cytoreductive efficacy of the individual components of multidrug chemotherapy for canine lymphoma is difficult to evaluate after complete remission. Objectives: To compare the cytoreductive efficacy of vincristine (VCR), cyclophosphamide (CPA), and doxorubicin (DXR) in dogs that received a 6-month modified version of the University of Wisconsin-Madison chemotherapy protocol (UW-25). Animals: Twenty-nine dogs with high-grade B-cell multicentric lymphoma. Methods: Rearranged immunoglobulin heavy chain gene fragments from lymphoma cells were amplified by polymerase chain reaction (PCR) and sequenced to prepare clone-specific primers and probes for real-time PCR. The number of lymphoma cells in peripheral blood was measured from diagnosis to week 11 of UW-25. Results: The number of lymphoma cells after the 1st administration of VCR, CPA, and DXR in weeks 1–4 was decreased in 29/29 (100%), 15/29 (51.7%), and 26/27 (96.3%) dogs, respectively. The cytoreductive efficacy of CPA was less than that of VCR and DXR. VCR, CPA, and DXR administered in weeks 6–9 were effective in 5/26 (19.2%), 5/20 (25.0%), and 14/19 (73.7%) dogs, respectively, indicating the sustained cytoreductive efficacy of DXR. CPA nonresponders were heavier and exhibited a shorter 1st remission than CPA responders. Conclusion and Clinical Importance: When using UW-25 for treatment of canine lymphoma, CPA was found to have less cytoreductive efficacy than VCR and DXR. Real-time PCR-based quantification of tumor cells is an objective marker of the efficacy of chemotherapeutic agents.

An efficient technique constructing anharmonic potential energy functions was applied to ab initio vibrational analysis of benzene. Anharmonic potentials including the 6-order terms, much higher than the full quartic force field, were automatically generated by a second-order algorithm using the scaled hypersphere search method, and vibrational calculations were performed at the level of VQDPT[1+2]. In comparison with previously reported anharmonic calculations, the present approach gave an excellent ab initio result for in-plane modes including the b 2u so called Kekule mode. Characteristic properties of various approaches were discussed in connection with the anharmonic effects and the strong vibronic effects.

The capacity to adsorb natural organic matter (NOM) and polystyrene sulfonates (PSSs) on small particle-size activated carbon (super-powdered activated carbon, SPAC) is higher than that on larger particle-size activated carbon (powdered-activated carbon, PAC). Increased adsorption capacity is likely attributable to the larger external surface area because the NOM and PSS molecules do not completely penetrate the adsorbent particle; they preferentially adsorb near the outer surface of the particle. In this study, we propose a new isotherm equation, the Shell Adsorption Model (SAM), to explain the higher adsorption capacity on smaller adsorbent particles and to describe quantitatively adsorption isotherms of activated carbons of different particle sizes: PAC and SPAC. The SAM was verified with the experimental data of PSS adsorption kinetics as well as equilibrium. SAM successfully characterized PSS adsorption isotherm data for SPACs and PAC simultaneously with the same model parameters. When SAM was incorporated into an adsorption kinetic model, kinetic decay curves for PSSs adsorbing onto activated carbons of different particle sizes could be simultaneously described with a single kinetics parameter value. On the other hand, when SAM was not incorporated into such an adsorption kinetic model and instead isotherms were described by the Freundlich model, the kinetic decay curves were not well described. The success of the SAM further supports the adsorption mechanism of PSSs preferentially adsorbing near the outer surface of activated carbon particles.

Reduction of oxide scale, which formed on low carbon steel sheet at 973K for 4.5 ks in 10.9 kPa CO2–14.5 kPa H2O–bal. N2, was studied at 673–1073K in pure H2. Reduction was slow at 673K and it became fast up to 823K. Between 823–873K, however, the reduction became slower than that at lower temperatures and showed a minimum at 873K. Then the reduction became faster with increasing temperature above 873K. The temperature at which the reduction was the slowest coincides with that of transformation between magnetite and wustite. Surface appearance of scales was different in these two temperature regions; surface cracks increased with reduction time at low temperature, whereas surface sintering proceeded with the time at high temperatures. The reason for the sudden reduction manner change around 873K was considered in terms of morphology, type and porosity of the oxide scales. It is concluded that the formar two can be the reason for the sudden change of reaction manner.

Options of systemic treatment for canine MCT have been still limited and most canine cases with MCTs eventually undergo relapses even after achievement of a remission. Thus additional therapies are required to establish for the tumor. To identify the novel candidate therapeutic targets for canine MCT, the mRNA expression and phosphorylation statuses of several receptor or non-receptor kinases as well as the inhibitory effect of 95 specific inhibitors on the growth were assessed in three canine MCT cell lines (HRMC, VIMC1 and CMMC1). Among the 14 targets, the mRNAs of 11, 7 and 7 kinases were amplified in HRMC, VIMC1 and CMMC1, respectively. The mRNAs of VEGFR3, PDGFRα, SRC, YES, LCK and FYN were detected in all cell lines. The phosphorylation of 12, 8 and 7 kinases was observed by using specific antibody arrays in HRMC, VIMC1 and CMMC1, respectively. DTK, EPHB6, AMPKα1, CREB, STAT5a and STAT5b were phosphorylated in all cell lines. The 10, 9 and 17 inhibitors exhibited the biological activity against the growth of HRMC, VIMC1 and CMMC1, respectively. Only three inhibitors such as SB218078 (for Chk1), PDGF RTK inhibitor IV (for PDGFR) and radicicol (for Hsp90) suppressed the growth of all three cell lines. The present study indicated that several kinases, such as Chk1, PDGFR and Hsp90, could be used as therapeutic targets in the treatment for canine MCT. Further studies and clinical trials are warranted to apply the inhibitors for the treatment of the tumor.

We retrospectively studied the clinical and laboratory features and outcomes of chronic enteropathy in Shiba dogs. Among 99 dogs with chronic enteropathy, 21 Shiba dogs (21%) were included in the study (odds ratio, 7.14). No significant differences were seen in signalment, clinical signs, symptoms or laboratory profiles between the Shiba and non-Shiba groups. Severe histopathological lesions in the duodenum were a common finding in the Shiba group. The median overall duration of survival in the Shiba group was 74 days, while that of the dogs in the non-Shiba group could not be determined because more than half of the cases remained alive at the end of this study. The difference between the groups was statistically significant (P

We evaluated the anti-tumor effect of adenoviral vector-mediated p53 gene therapy on the growth of canine osteosarcoma xenografts formed in nude mice. Nude mice were subcutaneously transplanted with cells of 2 P53 mutant canine osteosarcoma cell lines, POS and CHOS. The osteosarcoma xenografts were injected with either an adenoviral vector that expresses canine wild-type P53 (AxCA-cp53) or LacZ (AxCA-LacZ). Tumor growth was significantly inhibited in the xenografts injected with AxCA-cp53 in comparison to those injected with AxCA-LacZ or PBS during the observation period of 27 days. An increase of the amount of p21(WAF1/CDKN1A) mRNA, and the number of apoptotic cells was shown in the tumors injected with AxCA-cp53 in comparison to those injected with AxCA-LacZ or PBS. The present study revealed that the adenoviral vector-mediated p53 gene transfer had an anti-tumor effect in canine osteosarcoma xenografts formed in nude mice.

Huge numbers of structures, 7000 isomers and 26229 transition structures, have been searched for C6H6 in quantum chemical calculations by using a GRRM/SCC-DFTB program. The stability of benzene is ...

A direct reaction pathway for water–gas shift reaction (WGSR) in gas phase, H2O + CO → H2 + CO2, was discovered by using anharmonic downward distortion following method. The direct WGSR is accompan...

A 10-year-old American Shorthair cat with nasal discharge, anorexia, and weight loss was found to have pancytopenia and hyperproteinaemia. Bone marrow aspiration revealed atypical plasma cells that totalled 50% of the nucleated bone marrow cells. The number of atypical plasma cells progressively increased in the peripheral blood during the observation period of 64 days. The cat did not respond to treatments with melphalan, chlorambucil, and prednisolone, and died 71 days after the initial presentation. Clinical, cytological, histopathological, and immunohistochemical findings in this case supported the diagnosis of myeloma-related disorder (MRD) with leukaemic progression.

Geosmin and 2-methylisoborneol (2-MIB) are naturally occurring compounds responsible for musty-earthy-odors in surface water supplies. They are a severe problem confronting utilities worldwide. Adsorption by powdered activated carbon (PAC) is a widely used process to control this problem, but it has low efficiency, which engenders large budget spending for utilities services. Super-powdered activated carbon (S-PAC) is activated carbon with much finer particles than those of PAC. Experiments on geosmin and 2-MIB adsorptions on S-PAC and PAC were conducted. Geosmin and 2-MIB adsorption capacities on S-PAC were not smaller than those on PAC although natural organic matter, which adversely impacted the adsorption capacity of geosmin and 2-MIB, was more adsorbed on S-PAC than on PAC, meaning that the adsorption competition is less severe for S-PAC than for PAC.

The feasibility of in-line coagulation as a pretreatment for ceramic microfiltration (MF) was verified by comparing its efficiency in the removal of viruses with that of the traditional mechanical mixing approach for coagulation, and by examining the effect of coagulant dose and coagulation time on virus removal. The in-line coagulation–ceramic MF system efficiently removed bacteriophage Qβ and MS2: removal ratios were >8.2 log for infectious viruses and >5.4 log for total (infectious + inactivated) virus particles. These values were similar to those of the mechanical coagulation–ceramic MF system. The in-line coagulation system has potential as a useful pretreatment for the removal of viruses as an alternative to the mechanical mixing system, because the former efficiently removes viruses and has a smaller footprint in treatment plants. For the in-line coagulation–ceramic MF system, a coagulant dose of 1.08 mg-Al/L and a coagulation time of 1 min were required to achieve a high level of virus removal. Infectious Qβ and MS2 were removed to similar levels by the two precoagulation methods tested, but the removal of total MS2 particles was higher than that of Qβ particles, possibly because of the selective interaction with the cake layer.

The dynamics on the vacuum-ultraviolet one-photon ionization of a formamide-water cluster is investigated by a combination of theoretical reaction-path search and infrared spectroscopic methods. A keto-enol tautomerization of the formamide moiety occurs after photoionization by a catalytic action of the water molecule accompanied with its long-distance migration; the water molecule in the cluster migrates almost one turn around the formamide moiety. During the migration, the water molecule abstracts the proton of CH in the formamide moiety and carries it to the O atom side in the carbonyl group through a "catch and release"-type catalytic action.

Several studies indicated that KIT mutation could cause ligand-independent activation of c-Kit receptor in canine mast cell tumor (MCT). The objective of this study was to investigate mechanisms of c-Kit receptor activation in various canine MCT cell lines. Four cell lines, HRMC (derived from cutaneous MCT), VIMC1 (visceral MCT), CoMS1 (visceral MCT) and CMMC1 (cutaneous MCT), were cultured in stem cell factor (SCF, a ligand of c-Kit receptor)-free medium and subjected to analyses of KIT mutation, c-Kit receptor phosphorylation, SCF expression and the effects of SCF stimulation. In addition, the SCF/c-Kit receptor autocrine mechanism was verified in HRMC cells. HRMC cells expressed wild type c-Kit receptor. Both VIMC1 and CoMS1 cells had the same one amino acid (AA) substitution in the extracellular domain of c-Kit receptor. CMMC1 cells had at least three variants of c-Kit receptor such as one AA deletion in the extracellular domain (variant A), one AA substitution in the extracellular domain as well as an internal tandem duplication in the juxtamembrane domain (variant B), and a nonsense mutation (variant C). Both mature and immature forms of c-Kit receptor were observed and the c-Kit receptors were phosphorylated in all cell lines. While both mature and immature forms of c-Kit receptor were substantially phosphorylated in CMMC1 cells, the immature form was slightly phosphorylated in other cell lines. Phosphorylation of c-Kit receptor in HRMC, VIMC1 and CoMS1 cells were enhanced by SCF stimulation whereas no enhancement was observed in CMMC1 cells. There was no effect of SCF stimulation on proliferation of all the cell lines. SCF protein was detectable in only HRMC cells although mRNA expression of SCF was detected in all the cell lines. A tyrosine kinase inhibitor Dasatinib (internal inhibitor) inhibited c-Kit receptor phosphorylation in HRMC cells whereas anti-canine SCF antibody (external inhibitor) had no inhibitory effect. Thus there could be no external SCF/c-Kit receptor autocrine mechanism whereas there could be an internal autocrine mechanism within HRMC cells. The results indicated that consistent c-Kit receptor phosphorylation could be caused by the stimulation with autocrine SCF in HRMC cells while it could be caused by functional mutations of KIT in VIMC1, CoMS1 and CMMC1 cells. As the four canine MCT cell lines had various aberrations associated with c-Kit receptor phosphorylation, KIT mutation and SCF expression, such molecular biological diversity might reflect the different biological behavior in canine MCT.

Structural, energetic, and mechanistic features of the initial steps of the reaction of an O atom with Si(001)-(2 × 1) were modeled by the use of a Si9H12 + O system. Transition state structures (TSs) and equilibrium structures (EQs) as well as dissociation channels (DCs) were systematically searched using a global reaction route mapping (GRRM) technique on the potential energy surface (PES) based on the scaled hypersphere search (SHS) algorithm. The first six low-energy structures were consistent with known structure models for oxygen adsorption on the dimer of Si(001)-(2 × 1). Other structures with higher energies could be well interpreted as intermediates leading toward reaction paths for migration/dissociation occurring at high temperatures.

Structures of transition states (TSs) and minima on seam of crossing (MSXs) for potential energy surfaces (PESs) of acetone of the S0, S1, and T1 states were explored. On the basis of the results, we propose a new mechanism, slow intersystem crossing from S1 to T1 without seam of crossing, followed by CH3 dissociation via a TS on T1; this slow pathway will be overtaken by a more efficient S1 pathway for higher energy. This is consistent with the observed long lifetime of the S1 species. Moreover, four channels, including three new ones, were found to regenerate the ground state acetone from the S1 PES, and they all may be involved in the roaming channel that has been proposed recently as a new route of CO generation in a 230 nm photolysis. There are significant differences in MSX structures and energies between the present CASPT2 results and previous CASSCF results.

Background: Tumor cell burden in dogs with lymphoma cannot be assessed accurately by diagnostic evaluation during clinical complete remission (CR). Recent advances in polymerase chain reaction (PCR)-based methods enabled us to quantify minimal residual disease (MRD) in canine lymphoma. Hypothesis/Objectives: To quantify MRD in dogs with lymphoma treated with multidrug chemotherapy and to correlate it with remission duration after chemotherapy. Animals: Seventeen dogs with lymphoma that achieved CR by multidrug chemotherapy. Methods: Rearranged immunoglobulin heavy chain or T-cell receptor γ chain gene fragments from lymphoma cells were PCR amplified and sequenced to prepare clone-specific primers and probes for real-time PCR to quantify MRD. MRD in the peripheral blood was monitored during and at the end of a 25-week multidrug chemotherapy protocol. Correlation between MRD at the end of chemotherapy and remission duration after chemotherapy was analyzed. Results: MRD gradually decreased after initiation of multidrug chemotherapy, reached a nadir as low as

The effects of coagulant residuals on fouling of a nanofiltration (NF) membrane were investigated. Experiments were carried out with a laboratory-scale microfiltration (MF)–NF setup and a pilot MF–NF plant. In the laboratory-scale experiments, NF feed water was pretreated with poly-aluminum chloride (PACl) or alum followed by MF. NF membrane permeability declined when the feed water contained residual aluminum at 18 μg/L or more, but not when it was lower than 9 μg/L. When pretreated with ferric chloride, no substantial decline of NF membrane permeability was observed; residual iron did not affect the permeability. When SiO 2 was added to the water before the pretreatment with PACl, the NF membrane permeability declined at about double the speed. Thermodynamic calculations and elemental analysis of foulants recovered from the membranes indicated that the majority of inorganic foulants were compounds composed of aluminum, silicate, and possibly potassium. In the pilot plant, NF feed was pretreated by PACl. Transmembrane pressure for NF doubled over 4.5 months of operation. Although the aluminum concentration in the NF feed was not high (30 μg/L), analysis of membrane foulants revealed excessive accumulation of aluminum and silicate, also suggesting that aluminum residuals caused the membrane fouling by alumino-silicates or aluminum hydroxide.

Norovirus (NV) is an important human pathogen that causes epidemic acute nonbacterial gastroenteritis worldwide. Because of the lack of a cell culture system or an animal model for this virus, studies of drinking water treatment such as separation and disinfection processes are still hampered. We successfully estimated NV removal performance during a coagulation-rapid sand filtration process by using recombinant NV virus-like particles (rNV-VLPs) morphologically and antigenically similar to native NV. The behaviors of two widely accepted surrogates for pathogenic waterborne viruses, bacteriophages Qbeta and MS2, were also investigated for comparison with that of rNV-VLPs. Approximately 3-log(10) removals were observed for rNV-VLPs with a dose of 40 muM-Al or -Fe, as polyaluminum chloride at pH 6.8 or ferric chloride at pH 5.8, respectively. Smaller removal ratios were obtained with alum and ferric chloride at pH 6.8. The removal performance for MS2 was somewhat larger than that for rNV-VLPs, meaning that MS2 is not recommended as an appropriate surrogate for native NV. By comparison, the removal performance for Qbeta was similar to, or smaller than, that for rNV-VLPs. However, the removal performances for rNV-VLPs and Qbeta differed between the coagulation process and the following rapid sand filtration process. Therefore, Qbeta also is not recommended as an appropriate surrogate for native NV.

This study was conducted to map the acquired proviral insertions in the chromosomal genome of feline lymphoid tumors induced by feline leukemia virus (FeLV). Chromosome specimens of the lymphoid tumor-derived cell lines and normal cat lymphocytes were subjected to fluorescence in situ hybridization and tyramide signal amplification, using an exogenous FeLV-A genome as a probe. Specific hybridization signals were detected only on the metaphase chromosomes of the tumor cells. Poisson's distribution-based statistics indicated that 6 chromosomal loci in each cell line showed FeLV integration. In the examination of metaphase chromosomes of FL-74, FT-1 and KO-1 cells, significant signals were detected on B2p15-p14, B2q11, D1p14, E1p14-p13, E1q12 and F2q16; A2p23-p22, B2p15-p14, B4p15-p14, D4q23-q24, E1p14-p13 and E2p13-p12; and A2p22, A3q22, B1p13, B1q13, D1p13 and D3p15-p14, respectively. Consistently, Southern blot hybridization using an FeLV LTR-U3 probe specific for exogenous FeLV revealed the presence of at least 6 copies of exogenous FeLV proviruses at different integration sites in each cell line. These results indicate that there may be common FeLV integration sites at least in A2p22 and B2p15-p14. The cytogenetic analysis used in this study can promptly screen FeLV insertions and provide tags for identifying the novel common integration site.

The present study aimed to objectively evaluate the adverse events after the administration of chemotherapeutic agents used in the University of Wisconsin (UW)-Madison chemotherapy protocol (UW-25) for canine lymphoma, using the Veterinary Co-operative Oncology Group common terminology criteria for adverse events (VCOG-CTCAE). The medical records of 40 dogs with multicentric high-grade lymphoma that underwent UW-25 were reviewed. Gastrointestinal adverse events of grade 2 and above and blood/bone marrow adverse events of all grades were evaluated. Gastrointestinal adverse events occurring at least once during the entire period of UW-25 were observed in 50% (20/40), 17.9% (7/39), and 8.1% (3/37) of the dogs after the administration of vincristine (VCR), cyclophosphamide (CPA), and doxorubicin (DXR), respectively. Blood/bone marrow adverse events occurring at least once during UW-25 were observed in 57.5% (23/40), 41% (16/39), and 8.1% (3/37) of the dogs after the administration of VCR, CPA, and DXR, respectively. The rate of patients that experienced gastrointestinal adverse events was higher after the first administration of VCR than after the first administration of DXR. Findings obtained in this study will be helpful in predicting the adverse events that could occur when dogs with lymphoma are treated with UW-25.

症例は35歳，女性．健診の超音波検査で膵上縁に動脈瘤を指摘された．腹部血管造影検査で脾動脈起始部に径2cmの嚢状の動脈瘤を認め，破裂予防のためinterventional radiological treatment(IVR)を検討したが，瘤が大きく，流出入脾動脈の屈曲が強く，患者・家族へのinformed consentで小切開開腹となった．術前の動脈造影で瘤直前後の結紮で膵脾への側副血行による血流の温存が予想できた．動脈瘤への流入流出部で脾動脈をクランプし動脈瘤遠位側の膵脾への動脈血流維持を確認した後，脾動脈を結紮した．術後経過に問題なく，退院後の腹部dynamic CTでも瘤への血流はなく膵脾への動脈血流は保持されていた．脾動脈瘤はしばしば遭遇する疾患であるが，破裂する危険性があるため，瘤が発見された場合は何らかの治療が必要となる．多くはIVRが選択されるが，自験例のように瘤の形態や大きさ，存在部位などでIVRが困難である場合，開腹結紮術は一考されるべき治療である．

Purpose: In a few patients, cardiac perforation and aortic injury have occurred during the Nuss procedure for pectus excavatum. The article details a modification of this procedure that enables the prevention of fatal complications. Methods: Our subjects were 22 males and 13 females with pectus excavatum who were aged 8.2 ± 3.7 years. Their Haller's computed tomography index was 5.2 ± 1.5. An introducer is inserted into the pleura between the sternum and thymus instead of the thoracic depression under right thoracoscopic guidance. After the introducer reaches the internal cranial position of the left nipple, the thoracoscope is shifted to the left thoracic cavity. The introducer is subsequently guided to the left highest intercostal space under left thoracoscopic guidance. Results: A single bar was inserted in 34 patients; 2 bars were required in 1 patient. The operating time was 95 ± 27 minutes and blood loss was 11 ± 6 g. Cardiac perforation did not occur in any patient. Conclusions: Our modified technique has certain advantages: (1) the introducer does not rub against the pericardium and heart; (2) the tip of the introducer can be observed at all times with a thoracoscope; (3) the anterior mediastinum between the left and right thoracic cavities is very narrow at the cranial level; (4) the introducer can be accurately directed to the left highest intercostal space; and (5) hemostasis and no injury of the bilateral thoracic organs can be confirmed.

The quality of treated water produced by a drinking water treatment plant (DWTP) is affected by many factors. Relative impacts of various factors that affect the following treated water quality indices—turbidity, color, aluminum concentration and aggregate organic constituents—were investigated. Multivariate statistical analysis based on Hayashi's Quantification Theory Type 1 was performed on a dataset comprised of statistics on raw and treated water quality collected from thousands of DWTPs throughout Japan. Explanatory factors were the source of the raw water, water treatment process employed, plant capacity, total plant capacity of the water supplier (TCWS), raw water quality and pH of treated water. The statistical analyses mainly revealed that not only did obvious factors such as raw water quality affect treated water quality, but also more obscure factors such as plant capacity and TCWS. The results also imply that the larger the water supplier a DWTP of a given size belongs to, the higher the quality of treated water will be.

Conformationally dependent ionization of the simplest amino acid, glycine, is studied by Penning ionization electron spectroscopy with velocity-resolved metastable He*(2(3)S) atoms. The observed He I ultraviolet photoelectron and Penning ionization electron spectra are reproduced by superimposed theoretical spectra, assuming thermal distributions of conformers. The conformations of amino acids are determined by analyzing the observed Penning ionization cross sections, peak shifts, and collision energy dependences of partial ionization cross sections (CEDPICS). The Penning ionization cross sections are governed by collisionally accessible exterior electron densities. When the amino and carbonyl groups are exposed to He* access, the nonbonding orbitals of N (n(N)) and O atoms (n(O)) give rise to strong bands. The observed negative peak shifts and negative CEDPICS for the n(N) and n(O) orbitals suggest the presence of attractive interactions around their electron distributions. The most attractive wells are estimated to be approximately 400 meV in the direction of the n(N) orbitals by ab initio model calculations. A conformer possessing dual hydrogen bonds contributes predominantly to the spectra.

The global reaction route mapping (GRRM) method enabled an automated and a systematic search for routes of chemical reactions on a potential energy surface based on the anharmonic downward distortion following (ADDF) approach [Chem. Phys. Lett. 2004, 384, 277]. On the other hand, the microiteration technique [Mol. Phys. 2006, 104, 701] has been developed for full optimizations of transition state (TS) structures for reactions/transformations in large flexible molecular systems and successfully used in ONIOM(QM:MM) calculations. In the present paper, combining the GRRM method with the microiteration technique, we developed a microiteration-ADDF (μ-ADDF) method for automated and systematic TS exploration of large flexible molecular systems. We showed that the method works well with two test systems, (H2CO)(H2O)100 and Si6(C12H17)6, in the ONIOM(QM:MM) framework. It is noted that the present μ-ADDF method can be used for pure quantum mechanics (QM) or molecular mechanics (MM) systems (without ONIOM) and has been tested successfully in C6H10O pure QM calculations.

Outer valence ionic states of Cr(CO)(6) and (eta(5)-C(5)H(5))Co(CO)(2) were investigated by means of Penning ionization electron spectroscopy (PIES) upon collision with metastable He*(2(3)S) excited atoms as well as high-level ab initio molecular orbital calculations taking electron correlation effects into consideration. By the 2D measurement combining the collision-energy-resolved technique of the metastable atomic beam and electron spectroscopy, ionic-state-resolved measurement of collision energy dependence of partial ionization cross sections (CEDPICS) was carried out. Because partial Penning ionization cross sections can be connected to the spatial extent of corresponding molecular orbitals (MOs) outside the boundary surface of colliding He* atoms, different slopes of CEDPICS were related to anisotropic interaction around the MO region. The observed CEDPICS and PIES band intensity were consistent with the order of calculated ionic states.

Super-powdered activated carbon (S-PAC) is activated carbon of much finer particle size than powdered activated carbon (PAC). Geosmin is a naturally occurring taste and odor compound that impairs aesthetic quality in drinking water. Experiments on geosmin adsorption on S-PAC and PAC were conducted, and the results using adsorption kinetic models were analyzed. PAC pulverization, which produced the S-PAC, did not change geosmin adsorption capacity, and geosmin adsorption capacities did not differ between S-PAC and PAC. Geosmin adsorption kinetics, however, were much higher on S-PAC than on PAC. A solution to the branched pore kinetic model (BPKM) was developed, and experimental adsorption kinetic data were analyzed by BPKM and by a homogeneous surface diffusion model (HSDM). The HSDM describing the adsorption behavior of geosmin required different surface diffusivity values for S-PAC and PAC, which indicated a decrease in surface diffusivity apparently associated with activated carbon particle size. The BPKM, consisting of macropore diffusion followed by mass transfer from macropore to micropore, successfully described the batch adsorption kinetics on S-PAC and PAC with the same set of model parameter values, including surface diffusivity. The BPKM simulation clearly showed geosmin removal was improved as activated carbon particle size decreased. The simulation also implied that the rate-determining step in overall mass transfer shifted from intraparticle radial diffusion in macropores to local mass transfer from macropore to micropore. Sensitivity analysis showed that adsorptive removal of geosmin improved with decrease in activated carbon particle size down to 1microm, but further particle size reduction produced little improvement.

For the entry of lentivirus into target cells, fusion between its viral membrane and cellular membrane is essential. The present study was conducted to examine the inhibitory effect of modified peptides corresponding to heptad repeats (HR) 1 and 2 of feline immunodeficiency virus (FIV) envelope gp40 on the fusion between the viral and cellular membranes. FIV-N36 and FIV-C35 were synthesized as authentic peptides of the N-terminal HR1 domain and C-terminal HR2 domain of FIV gp40, respectively. FIV-C35EK1, FIV-C35EK2, and FIV-C35EK3 were peptides synthesized by modifying FIV-C35 as the X-EE-XX-KK concept to increase their solubility in water and the stability of their alpha-helicity. FIV-C35 and FIV-C35EK1 inhibited the cell membrane fusion mediated by FIV-infected cells and the replication of FIV. FIV-N36, FIV-C35EK2, and FIV-C35EK3 did not show any apparent inhibitory effect. These results indicated that the newly developed membrane fusion inhibitors could facilitate the development of novel anti-lentiviral chemotherapies.

Isomerization pathways of hexasilabenzene (Si6H6) were theoretically searched by using the anharmonic-downward-distortion-following (ADD-following) method, which is a recently established systematic and automatic transition-state (TS) finding method. The search revealed the lowest barrier from the six-membered-ring structure in hexasilabenzene, and the barrier height was computed to be only 74 kJ mol−1. It follows that the six-membered ring in hexasilabenzene can easily be deformed via the TS structure. The present study identified three lower-lying TS structures around hexasilabenzene by the systematic search. These TS structures will be useful for designing a suitable substituent in a hexasilabenzene derivative for its isolation in the future, since such a substituent should be designed so that all TSs deforming the six-membered-ring backbone in a hexasilabenzene derivative are destabilized.

A new proviral integration site for feline leukemia virus (FeLV), termed flit-1, was identified from feline thymic lymphoma. Among 35 FeLV-related tumors examined, 5 of 25 thymic lymphomas demonstrated proviral insertion within flit-1 locus whereas none of four alimentary and five multicentric lymphomas and one T-lymphoid leukemia examined had rearrangement in this region. Extensive sequence analysis has shown that flit-1, which is noncoding, is conserved on human chromosome 12 and mouse chromosome 15. The human and murine homologs of flit-1 are positioned approximately 30-kb upstream to activin-A receptor type II-like 1 (ACVRL1/ALK1) gene. Expression of ACVRL1 mRNA was examined in two of five lymphomas with flit-1 rearrangement and detected in both of the two whereas normal thymuses and seven lymphoid tumors without flit-1 rearrangement had no detectable expression. Therefore, flit-1 appears to represent a novel FeLV proviral common integration domain that may influence lymphomagenesis as insertional mutagenesis.

Time-course changes in the concentration of serum amyloid A (SAA), a major acute phase protein, were measured in a cat with pancreatitis over an 831-day period and compared with changes in WBC count and feline trypsin-like immunoreactivity (fTLI). SAA concentration was increased at the onset of the disease and gradually decreased over 5 days of treatment with an improvement in the clinical condition. In contrast, fTLI concentration and WBC count were not increased at the onset of the disease but increased gradually during the 5 days of treatment. Long-term monitoring from days 68 to 831 revealed a good correlation between SAA concentration and the reoccurrence of clinical signs in the cat; however, WBC count did not increase even with the exacerbation of disease. These findings suggest that the SAA concentration may be a useful marker for evaluating response to treatment and disease exacerbation in feline pancreatitis.

The reaction of formic acid with hydroxyl radical, which is considered to be relevant to atmospheric chemistry, has been extensively studied. A water-catalyzed process of this reaction is computationally studied here for the first time. The scaled hypersphere search method was used for global exploration of pre-reaction complexes. Calculations were performed at high level of theory, such as CCSD(T)/cc-pVTZ//B3LYP/6-311+G(2df, 2p) and CCSD(T)/cc-pVTZ//MP2/aug-cc-pVDZ. It is found that the water-catalyzed process of this reaction is more kinetically favorable than its non-catalytic process. Such catalytic process may also be of interest for atmospheric chemistry, like the non-catalytic one.

We have developed an apparatus for investigating isomer-selected photodissociation reactions by ion mobility spectrometry coupled with time-of-flight mass spectrometry. By using this apparatus we optimized ion intensities under isomer-separation condition for carbon cluster ions. Among ions having linear and cyclic isomers, we demonstrated dissociation of isomer-selected C9 + ion at 355-nm photon irradiation. As a result we observed marked difference in the dissociation pathways between from linear isomer and from cyclic isomer. For the purpose of adopting this technique to metal-ligand cluster ions, we also investigated mobility spectrometry of vanadium metal ions with Ar, in addition to electronic state separation for free atomic ions.