[Novel Treatment Strategy Using Trafermin® Consisting of bFGF for Intractable Pancreatic Fistula-A Case Report].

Pancreatic fistula is one of the most critical complication following distal pancreatectomy. We report here a successfully treated case with intractable pancreatic fistula using Trafermin® consisting of basic fibroblast growth factor(bFGF). A 60- year-old man underwent laparoscopic distal pancreatectomy. After surgery, pancreatic fistula was occurred. Pancreatic fistula persisted for 3 months despite of several conservative treatments. After obtaining informed consent, we started to inject 50µg/day of Trafermin® through a drainage tube into the dehiscence of pancreas. Consequently, pancreatic fistula was successfully closed within a week. This technique could be one of the treatment choices for intractable pancreatic fistula following distal pancreatectomy.

[Mediastinoscopic Radical Esophagectomy for Esophageal Cancer in a Patient with Chronic Pulmonary Infection].

Mediastinoscopic esophagectomy(ME)is a minimally invasive approach without thoracotomy and pulmonary atelectasis during surgery. Here, we report the case of a 67-year-old patient who was successfully treated with thoracic esophageal cancer and severe chronic pulmonary infection using ME and home enteral nutrition therapy. Esophageal cancer patients with severe lung dysfunction have a risk of postoperative pneumoniae. ME could be a promising procedure for patients with severe lung dysfunction. We highlight the usefulness of ME as a safe approach to avoid pulmonary complications.

[A Case of G-CSF Producing Esophageal Carcinosarcoma Treated Effectively with Nivolumab for Recurrences following Two-Stage Surgery].

G-CSF producing esophageal carcinosarcoma is extremely rare, and its effective treatment strategy remains undefined. Here, we report the case of a 69-year-old woman who underwent successful two-stage surgery using mediastinoscopic esophagectomy and laparoscopic reconstruction for the management of severe anemia, malnutrition, and inflammation due to G-CSF producing esophageal carcinosarcoma(G-CSF 265 pg/mL). Chemoradiotherapy could not manage lymph node recurrences in the patient; however, nivolumab was found to be effective and helped achieved a prolonged partial response.

[Conversion Surgery for Advanced Gastric Cancer with Ovarian Metastasis-A Case Report with Review of the Literatures].

Stage Ⅳ gastric cancer(GC)with ovarian metastasis showed poor prognosis and its treatment strategy remains unclear. Recent studies identified the favorable prognostic effect of conversion surgery in Stage Ⅳ GC following intensive chemotherapy. We report here a case with advanced GC and ovarian metastasis, who underwent conversion surgery for them followed by chemotherapy and had a long-term survival. We reviewed the literatures in order to discuss clinical significance of our treatment strategy.

Efficient and rapid assessment of multiple aspects of frailty using the Kyoto Frailty Scale, developed from the Edmonton Frail Scale.

[Purpose] Global aging has led to a dramatic increase in the number of frail people, who are likely to become bedridden. Since frailty can be partially reversed, early intervention would be beneficial for patients, family members, and clinicians. This study was designed to develop a screening tool for an accurate and comprehensive assessment of frailty by modulating the Edmonton Frail Scale (EFS). [Participants and Methods] The EFS, covering multiple domains, is one of the major diagnostic tools for frailty. Frail and non-frail participants (n=67) were evaluated for each diagnostic item of the EFS to identify the most efficient combination of questions by evaluating its sensitivity and specificity. [Results] The Kyoto Frailty Scale (KFS) was developed as a rapid frailty scale, based on the EFS. The KFS comprises nine questions about health status, polypharmacy, hospitalization, living with a reliable caregiver, shopping, transportation, housework, money management, and forgetting to take medicine. The KFS has an excellent negative predictive value (100%) for screening frailty and a positive predictive value (97%) for screening prefrailty and frailty if we regard KFS ≥4 as a test positive. [Conclusion] The KFS permits clinician to rapidly and accurately screen for frailty and prefrailty, or exclude frailty.

[Better Effect of Home Night Enteral Nutrition in an Elderly Patient Following Laparoscopic Total Gastrectomy for Advanced Gastric Cancer].

Postoperative low nutrition, body weight loss and poor QOL are pivotal clinical issues for elderly patients following total gastrectomy for advanced gastric cancer. Here, we report a successfully treated high-risk elderly case using home night enteral nutrition and adjuvant chemotherapy following laparoscopic total gastrectomy for advanced gastric cancer. Home night enteral nutrition might not impair oral intake and daily QOL, and could be a useful treatment strategy for elderly patients following total gastrectomy.

[Long-Term Survivor with Recurrent Gastric Cancer Using Trifluridine/Tipiracil as a Late-Line Chemotherapy].

TAGS trial revealed the efficacy and safety of trifluridine/tipiracil(Lonsurf®)treatment in patients with metastatic gastric cancer following gastrectomy. Here, we successfully treated 38 months survival case after recurrences following radical gastrectomy for advanced adenocarcinoma of esophago-gastric junction using historical recommended chemotherapy regimens and trifluridine/tipiracil as a fifth-line chemotherapy. Trifluridine/tipiracil therapy contributed to effective and safety treatment even in late-line chemotherapy for recurrent gastric cancer.

[A Case of Peritoneal Metastasis from an Unresectable Advanced Gastric Cancer with a Good Response to Nivolumab].

A 74-year-old woman presented with abdominal discomfort and was diagnosed with an unresectable advanced gastric cancer(T4aN3aM1, stage Ⅳ)based on a thorough examination. S-1/cisplatin therapy was administered as first-line treatment and paclitaxel/ramucirumab therapy as the second-line treatment. However, because the patient developed a peritoneal dissemination and her lymph node metastasis increased despite these regimens, nivolumab was introduced as a third- line treatment. The CT scan revealed that after the 5 courses of nivolumab, both the peritoneal dissemination and metastatic lymph nodes shrunk; after 12 courses of nivolumab, the peritoneal dissemination almost disappeared. Although nivolumab, an anti-programmed cell death-1(PD-1)antibody, has the possibility to cause immune-related adverse events not seen with conventional chemotherapy, in the present case, these events did not occur and the antitumor effects were maintained for a relatively long period without a decrease in the performance status(PS). We experienced a case of peritoneal metastasis from gastric cancer with a good response to nivolumab. Herein, this case is reported with some literature review.

Crystal structure of tRNA m(1)A58 methyltransferase TrmI from Aquifex aeolicus in complex with S-adenosyl-L-methionine.

The N (1)-methyladenosine residue at position 58 of tRNA is found in the three domains of life, and contributes to the stability of the three-dimensional L-shaped tRNA structure. In thermophilic bacteria, this modification is important for thermal adaptation, and is catalyzed by the tRNA m(1)A58 methyltransferase TrmI, using S-adenosyl-L-methionine (AdoMet) as the methyl donor. We present the 2.2 Å crystal structure of TrmI from the extremely thermophilic bacterium Aquifex aeolicus, in complex with AdoMet. There are four molecules per asymmetric unit, and they form a tetramer. Based on a comparison of the AdoMet binding mode of A. aeolicus TrmI to those of the Thermus thermophilus and Pyrococcus abyssi TrmIs, we discuss their similarities and differences. Although the binding modes to the N6 amino group of the adenine moiety of AdoMet are similar, using the side chains of acidic residues as well as hydrogen bonds, the positions of the amino acid residues involved in binding are diverse among the TrmIs from A. aeolicus, T. thermophilus, and P. abyssi.

Extracellular glypican-1 affects tumor progression and prognosis in esophageal cancer.

INTRODUCTION: Cells are covered with a glycan surface layer that is referred to as the glycocalyx (GCX). It has been reported that the formation of the GCX is promoted on cancer cells and is associated with tumor growth and metastasis. Heparan sulfate proteoglycan glypican-1 (GPC1) is a core protein of the GCX that is overexpressed in esophageal squamous cell carcinoma (ESCC) and is involved in the development and progression of cancer cells. The purpose of the present study is to analyze the utility of GPC1 as a new biomarker ralated to glycocalyx that reflects therapeutic effect and prognosis of ESCC. METHODS: We measured the concentration of GPC1 protein in preoperative plasma from advanced esophageal cancer patients and examined its relationships with clinicopathological factors and therapeutic efficacy, and the effects of extracellular GPC1 were investigated. RESULTS: The following clinical factors were significantly correlated with the preoperative high GPC1 concentration: male, tumor size ≥30 mm, venous invasion, pT factor ≥2, pStage ≥3, residual tumor, and distant metastatic recurrence. Both overall and recurrence-free survival were significantly worse in the high GPC1 group. Extracellular GPC1 protein concentration reflected intracellular GPC1 expression. Furthermore, we examined the effects of extracellular recombinant human (rh)GPC1 on ESCC cells, and found that extracellular rhGPC1 affects cell motility, including migration and invasion. CONCLUSIONS: These results demonstrated the utility of extracellular GPC1 as a biomarker, which can be assayed from a less invasive blood sample-based liquid biopsy. Extracellular GPC1 protein plays a role in both tumor cell motility and cancer progression. Thus, plasma GPC1 is a useful biomarker for esophageal cancer progression and may be a potential candidate of therapeutic target.

Characterization and structure of the Aquifex aeolicus protein DUF752: a bacterial tRNA-methyltransferase (MnmC2) functioning without the usually fused oxidase domain (MnmC1).

Post-transcriptional modifications of the wobble uridine (U34) of tRNAs play a critical role in reading NNA/G codons belonging to split codon boxes. In a subset of Escherichia coli tRNA, this wobble uridine is modified to 5-methylaminomethyluridine (mnm(5)U34) through sequential enzymatic reactions. Uridine 34 is first converted to 5-carboxymethylaminomethyluridine (cmnm(5)U34) by the MnmE-MnmG enzyme complex. The cmnm(5)U34 is further modified to mnm(5)U by the bifunctional MnmC protein. In the first reaction, the FAD-dependent oxidase domain (MnmC1) converts cmnm(5)U into 5-aminomethyluridine (nm(5)U34), and this reaction is immediately followed by the methylation of the free amino group into mnm(5)U34 by the S-adenosylmethionine-dependent domain (MnmC2). Aquifex aeolicus lacks a bifunctional MnmC protein fusion and instead encodes the Rossmann-fold protein DUF752, which is homologous to the methyltransferase MnmC2 domain of Escherichia coli MnmC (26% identity). Here, we determined the crystal structure of the A. aeolicus DUF752 protein at 2.5 Å resolution, which revealed that it catalyzes the S-adenosylmethionine-dependent methylation of nm(5)U in vitro, to form mnm(5)U34 in tRNA. We also showed that naturally occurring tRNA from A. aeolicus contains the 5-mnm group attached to the C5 atom of U34. Taken together, these results support the recent proposal of an alternative MnmC1-independent shortcut pathway for producing mnm(5)U34 in tRNAs.

The Role of Inflammation-associated microRNA-4257 as a Promoter of Malignancy in Colorectal Cancer.

BACKGROUND/AIM: The clinical significance ofmiR-4257 in patients with colorectal cancer (CRC) remains unclear. Here, we investigated the usefulness of measuring miR-4257 levels in the plasma and cancer tissues of patients with CRC, and the function of miR-4257 in CRC cell lines. MATERIALS AND METHODS: miR-4257 levels were measured in the plasma and cancer tissues of patients with CRC using quantitative polymerase chain reaction. The relationships between miR-4257 level and clinicopathological features were examined. Proliferation, transwell, wound healing, and adhesion assays were performed using a miR-4257 mimic and inflammatory cytokines. RESULTS: Relapse-free survival was significantly lower in patients with high miR-4257 levels in the plasma and cancer tissue (p<0.001 andp=0.016, respectively). High miR-4257 expression was an independent predictive factor for recurrence (p=0.017 and p=0.028). Addition of inflammatory cytokines to CRC and normal cell lines increased the expression of miR-4257 in the cell lines and cell culture medium. Over-expression of miR-4257 in CRC cells increased malignancy, while over-expression in normal cells increased adhesion to CRC cells. The addition of inflammatory cytokines to normal cell lines enhanced adhesion to CRC cell lines. CONCLUSION: miR-4257 level in plasma and cancer tissues is a biomarker of disease recurrence in patients with CRC. Moreover, miR-4257 promoted tumour growth and was associated with cancer-induced inflammation.

Core conversion reactions of the cubane-type metal-sulfido clusters: shape shift, contraction, and expansion of the MM'Re2S4 Cubanes (M = Ir, Rh, Ru; M' = Pt, Pd).

Treatment of incomplete cubane-type clusters [(Cp*M){Re(L)}(2)(mu(3)-S)(mu(2)-S)(3)] (M = Ir (1a), Rh (1b); Cp* = eta(5)-C(5)Me(5); L = S(2)C(2)(SiMe(3))(2)) and [{(Pmb)Ru}{Re(L)}(2)(mu(3)-S)(mu(2)-S)(3)] (Pmb = eta(6)-C(6)Me(5)H) with 1 equiv of [Pt(PPh(3))(3)] gave tetranuclear tetra(sulfido) clusters having raft-type cores, [(Cp*M){Pt(PPh(3))(2)}{Re(L)}(2)(mu(3)-S)(4)] (M = Ir (3a), Rh) and [{(Pmb)Ru}{Pt(PPh(3))(2)}{Re(L)}(2)(mu(3)-S)(4)], which presents a sharp contrast to the reactions with [Pd(PPh(3))(4)] reported previously, affording the cubane-type clusters [(Cp*M){Pd(PPh(3))}{Re(L)}(2)(mu(3)-S)(4)] (M = Ir (2a), Rh) and [{(Pmb)Ru}{Pd(PPh(3))}{Re(L)}(2)(mu(3)-S)(4)]. The reactions of 2a with diphosphines P2 resulted in the conversion of its cubane-type core into the analogous raft-type frameworks, forming [(Cp*Ir){Pd(P2)}{Re(L)}(2)(mu(3)-S)(4)] (P2 = cis-Ph(2)PCH=CHPPh(2) (6), Ph(2)PCH(2)CH(2)PPh(2), Ph(2)PCH(2)CH(2)CH(2)PPh(2)). On the other hand, when 2 was allowed to react with Ph(2)PCH(2)PPh(2) (dppm) as P2, the trinuclear tri(sulfido) cluster [(Cp*Ir){Re(L)}(2)(mu(3)-S)(2)(mu(2)-S)(mu(2)-dppm)] (9a) was obtained. Alternatively, this cluster 9a and its Rh analogue 9b were derived from the incomplete cubane-type clusters 1a and 1b by treatment with dppm. It has also been found that further treatment of the cubane-type cluster 2a with excess [Pd(PPh(3))(4)] affords the heptanuclear tetra(sulfido) cluster [(Cp*Ir){Pd(PPh(3))}(4)Re(2)(mu(3)-L)(2)(mu(3)-S)(4)] (10). The detailed structures have been determined by the X-ray analyses for 3a, 6, 9a, and 10.

Structural basis of the initial binding of tRNA(Ile) lysidine synthetase TilS with ATP and L-lysine.

In the bacterial genetic-code system, the codon AUA is decoded as isoleucine by tRNA(Ile)(2) with the lysidine residue at the wobble position. Lysidine is derived from cytidine, with ATP and L-lysine, by tRNA(Ile) lysidine synthetase (TilS), which is an N-type ATP pyrophosphatase. In this study, we determined the crystal structure of Aquifex aeolicus TilS, complexed with ATP, Mg2+, and L-lysine, at 2.5 A resolution. The presence of the TilS-specific subdomain causes the active site to have two separate gateways, a large hole and a narrow tunnel on the opposite side. ATP is bound inside the hole, and L-lysine is bound at the entrance of the tunnel. The conserved Asp36 in the PP-motif coordinates Mg2+. In these initial binding modes, the ATP, Mg2+, and L-lysine are held far apart from each other, but they seem to be brought together for the reaction upon cytidine binding, with putative structural changes of the complex.

Overexpression of cyclase-associated protein 2 in multistage hepatocarcinogenesis.

PURPOSE: Hepatocellular carcinoma (HCC) associated with chronic liver disease is known to show an obvious multistage process of tumor progression. We previously identified heat shock protein 70 as a molecular marker of early HCC during investigation of expression profiling in multistage hepatocarcinogenesis. In this report, we examined cyclase-associated protein 2 (CAP2), which is also listed as an up-regulated gene in early HCC. EXPERIMENTAL DESIGN: We measured the level of CAP2 mRNA by real-time quantitative PCR. We raised a polyclonal antibody against CAP2 and we confirmed the expression of CAP2 by immunoblotting and immunohistochemistry in HCC cell lines and HCC tissues. RESULTS: According to real-time quantitative PCR, the level of CAP2 mRNA was up-regulated in early HCC when compared with noncancerous liver tissue, and it was further up-regulated in progressed HCC. We raised a polyclonal antibody against CAP2, which showed a single 53-kDa band of strong intensity in the human HCC cell lines and HCC tissues but only a weak band in the noncancerous liver tissues in Western blot analysis. Immunohistochemical examination of CAP2 revealed its significant overexpression in early HCC when compared with noncancerous and precancerous lesions and in progressed HCC when compared with early HCC. CONCLUSION: Our findings show that CAP2 is up-regulated in HCC when compared with noncancerous and precancerous lesions. This is the first report that proves that CAP2 is up-regulated in human cancers and that this is possibly related to multistage hepatocarcinogenesis.

Establishment of perineural invasion models and analysis of gene expression revealed an invariant chain (CD74) as a possible molecule involved in perineural invasion in pancreatic cancer.

PURPOSE: Perineural invasion causes frequent local recurrence even after resection and a poor prognosis for pancreatic cancer. We established perineural invasion models and analyzed the molecular mechanism of perineural invasion in pancreatic cancer. EXPERIMENTAL DESIGN: Seven pancreatic cancer cell lines with or without human peripheral nerves were s.c. implanted in nonobese diabetes/severe combined immunodeficient mice. We compared expression profiles among high and low perineural invasion cell lines by using an oligonucleotide microarray. We examined up-regulation of the invariant chain (CD74) in high perineural invasion cell lines in mRNA and protein levels and surgical cases immunohistochemically. RESULTS: Four of seven pancreatic cancer cell lines (CaPan1, CaPan2, CFPAC, and MPanc96) showed perineural invasion to s.c. transplanted human peripheral nerves. Moreover, CaPan1 and CaPan2 (high perineural invasion group) also resulted in a high frequency of perineural invasion to mouse s.c. peripheral nerves, whereas three pancreatic cancer cell lines HPAFII, AsPC1, and Panc1 (low perineural invasion group) did not show perineural invasion to either human or mouse nerves. We identified 37 up-regulated genes and 12 down-regulated genes in the high perineural invasion group compared with the low perineural invasion group. Among them, CD74 was up-regulated in the high perineural invasion group in mRNA and protein levels. Furthermore, immunohistochemical expression of CD74 in clinical cases revealed its significant overexpression in pancreatic cancer with perineural invasion (P < 0.008). CONCLUSIONS: This is the first report of perineural invasion models using human pancreatic cancer cell lines. In combination with gene expression profiling, it was indicated that CD74 could be a candidate molecule involved in perineural invasion. These models provide new approaches for study of perineural invasion in pancreatic cancer.

Crystallization of the archaeal transcription termination factor NusA: a significant decrease in twinning under microgravity conditions.

The transcription termination factor NusA from Aeropyrum pernix was crystallized using a counter-diffusion technique in both terrestrial and microgravity environments. Crystallization under microgravity conditions significantly reduced the twinning content (1.0%) compared with terrestrially grown crystals (18.3%) and improved the maximum resolution from 3.0 to 2.29 A, with similar unit-cell parameters. Based on a comparison of the crystal parameters, the effect of microgravity on protein crystallization is discussed.

[Bacteriological analysis of the risk of contamination in the catheter package for intravenous hyperalimentation--the frequency of catheter package exchange for IVH].

The all-in-one catheter package for intravenous hyperalimentation provided an easy handling to the medical staff. However, the package is the cause for a medical cost increase. In order to reduce the medical cost, the incidence of bacterial contamination on the catheter was compared between the two groups: one group is to exchange the catheter once a week and the other is to exchange the catheter twice a week. No bacterial contamination was found in both groups. The result suggested that once a week catheter exchange was tolerable against bacterial contamination.

Crystal structure of tRNA pseudouridine synthase TruA from Thermus thermophilus HB8.

The pseudouridine synthase (Psi synthase) TruA catalyzes the conversion of uridine to pseudouridine at positions 38, 39 and/or 40 in the anticodon stem-loop (ASL) of tRNA. We have determined the crystal structure of TruA from Thermus thermophilus HB8 at 2.25 A resolution. TruA and the other (Psi synthases have a completely conserved active site aspartate, which suggests that the members of this enzyme family share a common catalytic mechanism. The T. thermophilus TruA structure reveals the remarkably flexible structural features in the tRNA-binding cleft, which may be responsible for the primary tRNA interaction. In addition, the charged residues occupying the intermediate positions in the cleft may lead the tRNA to the active site for catalysis. Based on the TruB-tRNA complex structure, the T. thermophilus TruA structure reveals that the tRNA probably makes the melting base pairs move into the cleft, and suggests that a conformational change of the substrate tRNA is necessary to facilitate access to the active site aspartate residue, deep within the cleft.

Crystal structure of a predicted phosphoribosyltransferase (TT1426) from Thermus thermophilus HB8 at 2.01 A resolution.

TT1426, from Thermus thermophilus HB8, is a conserved hypothetical protein with a predicted phosphoribosyltransferase (PRTase) domain, as revealed by a Pfam database search. The 2.01 A crystal structure of TT1426 has been determined by the multiwavelength anomalous dispersion (MAD) method. TT1426 comprises a core domain consisting of a central five-stranded beta sheet surrounded by four alpha-helices, and a subdomain in the C terminus. The core domain structure resembles those of the type I PRTase family proteins, although a significant structural difference exists in an inserted 43-residue region. The C-terminal subdomain corresponds to the "hood," which contains a substrate-binding site in the type I PRTases. The hood structure of TT1426 differs from those of the other type I PRTases, suggesting the possibility that TT1426 binds an unknown substrate. The structure-based sequence alignment provides clues about the amino acid residues involved in catalysis and substrate binding.