Identification of genes responsible for absorbing palladium ion in Escherichia coli.

The capability of Escherichia coli BW25113 to adsorb palladium (Pd) ions in a single-gene-knockout library was investigated using high-throughput screening. The results revealed that compared to BW25113, nine strains promoted Pd ion adsorption, whereas 22 strains repressed. Although further studies are required because of the first screening results, our results will provide a new perspective for improving the biosorption.

[A Case of paraganglioma of Urinary Bladder].

A 66-year-old woman was referred to our hospital because a bladder tumor was detected by abdominal ultrasonography. Although she was asymptomatic, cystoscopy showed the nodular sessile tumor in the bladder. We performed transurethral resection of the bladder tumor (TURBT) and histological examination revealed paraganglioma of the urinary bladder. Computed tomography (CT) and metaiodobenzylguanidine (MIBG) scintigraphy did not reveal any other lesions of paraganglioma. One month later, we performed a second TURBT, but the histological examination revealed no residual tumor. She has been followed up for 3 years after operation without any evidence of recurrence. Paraganglioma of the urinary bladder originates from chromaffin tissue of the sympathetic nervous system associated with the urinary bladder wall. For bladder submucosal tumor, we should consider bladder paraganglioma. And second TURBT is a useful option for the evaluation of the residual tumor.

Thioredoxin, an antioxidant redox protein, in ovarian follicles of women undergoing in vitro fertilization.

Oxidative stress has a bidirectional role in the development and maturation of zygotes and embryos. Reduction-oxidation reactions and regulatory proteins, such as thioredoxin (TRX) and thioredoxin reductase (TRXR), are intimately involved in the regulation of oxidative stress. The aim of this study was to determine the levels of TRX mRNA and protein in ovarian follicles collected from women undergoing in vitro fertilization (IVF) and to assess these levels relative to follicle size, presence of oocytes, and responsiveness to superovulation. Follicular fluid (FF) and/or granulosa cells (GCs) from large and small follicles were collected at the time of ovum pick-up from 42 IVF patients enrolled in this study. We divided the patients into normal and poor responders (NR and PR, respectively) based on the serum estradiol levels on the day of human chorionic gonadotropin (hCG) administration. We also compared the TRX concentration in FF (FF-TRX) between oocyte-containing follicles (Oc+) and empty follicles (Oc-). The transcript levels of TRX, but not TRXR, were significantly higher in GCs derived from follicles collected from NR than PR, as determined by semi-quantitative RT-PCR analysis. In NR, the FF-TRX was significantly higher in Oc+ follicles than in Oc- follicles and also in large Oc+ follicles than in large Oc- follicles. Unlike NR, PR exhibited no positive association with elevated FF-TRX and presence of oocytes. Based on its collective anti-oxidative, cytoprotective, and cytokine-like properties of TRX, TRX is likely to be involved in the optimal growth and maturation of ovarian follicles and responsiveness to hyperstimulation.

A case of adenosquamous carcinoma of the lower bile duct diagnosed preoperatively via transpapillary biopsy.

A 78-year-old man presented to our hospital with fever and brownish urine. Upon thorough examination, a diagnosis of obstructive jaundice and acute cholangitis associated with a lower bile duct tumor was made. Endoscopic retrograde cholangiopancreatography revealed entire circumferential stenosis of the lower bile duct. Examination of a transpapillary biopsy specimen of the lesion suggested adenosquamous carcinoma. The patient underwent subtotal stomach-preserving pancreaticoduodenectomy. Histopathological examination revealed adenocarcinoma of the lower bile duct and squamous cell carcinoma components;a case of adenosquamous carcinoma was accordingly diagnosed. The lower bile duct tumor directly extended into the pancreatic parenchyma for approximately 1mm. We performed radical surgery and administered adjuvant chemotherapy with gemcitabine because of advanced neural invasion after consulting with the patient. There was no sign of recurrence 46 months after surgery. As adenosquamous carcinoma of the extrahepatic bile duct is rare, it is difficult to preoperatively diagnose the condition. Only a few cases have been reported till date.

[mFOLFOX6 chemotherapy enabling curative resection in a patient with locally advanced duodenal adenocarcinoma and lymph node metastases].

A 65-year-old man was referred to our hospital with a primary complaint of unintentional weight loss. Upper gastrointestinal endoscopy showed a large duodenal tumor, and computed tomography (CT) showed tumor invasion of the pancreatic head, associated with lymph node swelling around the tumor and abdominal aorta. After 4 courses of chemotherapy with 5-fluorouracil, leucovorin, and oxaliplatin (mFOLFOX6), follow-up upper gastrointestinal endoscopy and biopsy showed no evidence of a duodenal tumor. However, follow-up CT showed persistent lymph node swelling near the superior mesenteric artery. The patient therefore underwent pancreaticoduodenectomy with curative intent. At 18 months after the surgery, there were no findings suggestive of tumor recurrence.

[A case of hypercalcemia associated with parathyroid hormone-related protein produced by the recurrence of B-cell lymphoma of the pancreas].

An 87-year-old woman was diagnosed with primary diffuse large B-cell lymphoma of the pancreas by endoscopic ultrasonography-guided fine needle aspiration. Complete remission was achieved after treatment with six courses of R-CHOP chemotherapy. However, two and a half years later, she was readmitted because of weakness during walking. At this time, laboratory tests revealed hypercalcemia associated with high plasma levels of parathyroid hormone-related protein (PTHrP), but bone lesions were not detected. Although computed tomography only revealed splenomegaly, we suspected a recurrence of her malignant lymphoma because she also had marked elevation of soluble interleukin-2 receptor and lactate dehydrogenase levels. Bone marrow examination revealed the involvement of Burkitt's lymphoma cells with malignant transformation. Immunohistochemical analysis confirmed that hypercalcemia was caused by a paraneoplastic syndrome related to PTHrP-producing B-cell lymphoma cells. Unfortunately, the patient's general condition rapidly deteriorated, and she died soon after admission. Our case is unusual because of the presentation of bone marrow relapse of malignant lymphoma.

A case of carcinosarcoma of the duodenum.

A 59-year-old man was admitted to the hospital because of upper abdominal pain. Endoscopic examination and computed tomography showed a polypoid tumor located in the 2nd portion of the duodenum with invasion to the pancreas head, and biopsy findings suggested a gastrointestinal stromal tumor. He underwent a pancreaticoduodenectomy, and the tumor, which measured 6.5×6.5 cm, was resected. Histologically, the tumor contained two divergent components: differentiated tubular adenocarcinoma and sarcomatoid tissue composed of spindle tumor cells. The tumor directly extended to the pancreas head and metastasized to multiple lymph nodes. The adenocarcinoma cells were positive for AE1/3 and cytokeratin 7 and negative for vimentin. In contrast, the sarcomatoid tissue was negative for epithelial markers and positive for vimentin. The tumor was finally diagnosed as duodenal carcinosarcoma. Duodenal carcinosarcoma is very rare, and only seven cases have been reported to date.

Exposure of the yeast Saccharomyces cerevisiae to functionalized polystyrene latex nanoparticles: influence of surface charge on toxicity.

Novel nanoparticles with unique physicochemical characteristics are being developed with increasing frequency, leading to higher probability of nanoparticle release and environmental accumulation. Therefore, it is important to assess the potential environmental and biological adverse effects of nanoparticles. In this study, we investigated the toxicity and behavior of surface-functionalized nanoparticles toward yeast (Saccharomyces cerevisiae). The colony count method and confocal microscopy were used to examine the cytotoxicity of manufactured polystyrene latex (PSL) nanoparticles with various functional groups (amine, carboxyl, sulfate, and nonmodified). S. cerevisiae were exposed to PSL nanoparticles (40 mg/L) dispersed in 5-154 mM NaCl solutions for 1 h. Negatively charged nanoparticles had little or no toxic effect. Interestingly, nanoparticles with positively charged amine groups (p-Amine) were not toxic in 154 mM NaCl, but highly toxic in 5 mM NaCl. Confocal microscopy indicated that in 154 mM NaCl, the p-Amine nanoparticles were internalized by endocytosis, whereas in 5 mM NaCl they covered the dead cell surfaces. This demonstrates that nanoparticle-induced cell death might to be related to their adhesion to cells rather than their internalization. Together, these findings identify important factors in determining nanoparticle toxicity that might affect their impact on the environment and human health.

[Intrascrotal malignant mesothelioma diagnosed after surgery for hydrocele testis: a case report].

We report here a case of intrascrotal malignant mesothelioma, arising from the tunica vaginalis, which was diagnosed after surgery for hydrocele testis. A 52-year-old man underwent left hydrocelectomy for hydrocele testis. After pathological diagnosis as malignant mesothelioma from the specimen of tunica vaginalis, left radical orchiectomy was performed. The patient had no exposure to asbestos and there has been no evidence of recurrence.

Molecular analysis of gastric differentiated-type intramucosal and submucosal cancers.

Identification of the molecular characteristics of intramucosal (IMCs) and submucosal cancers (SMCs) is essential to our understanding of early gastric carcinogenesis. However, little is known regarding the differences between the 2 lesions. One hundred and forty-eight patients with primary early gastric cancer [IMC, 106; SMC, 42] were characterized for expression of cell cycle-related proteins and loss of heterozygosity (LOH). We also examined microsatellite instability (MSI) and methylation status. For LOH and methylation studies, we used a panel of 17 microsatellite markers (3p, 4p, 5q, 9p. 13q, 17p, 18q and 22q) and promoter regions of 9 genes (MLH-1, RUNX3, p16, HPP1, RASSF2A, SFRP1, DKK-1, ZFP64 and SALL4) that are frequently altered or methylated in gastric cancers. Overexpression of p53 and cyclin D1 was observed in SMC. In addition, low expression of p27 was more frequent in SMC than in IMC. Frequencies of 4p, 9p, 13q and 22q were significantly higher in SMC than in IMC. The SALL4 gene was frequently methylated in SMC compared with IMC. However, other gene methylations were common in both IMC and SMC. The frequency of LOH-high status/methylation-low status was significantly higher in SMC than in IMC. However, LOH-low status/methylation-high status in SMC was more frequently found in IMC. Our data confirm that methylation of cancer-related genes plays a major role in the development of IMCs. Importantly, the results also show that gastric submucosal progression is characterized by the accumulation of specific genetic alterations. In addition, changes of cell cycle-related proteins are associated with cancer progression.

[A case of extramedullary plasmacytoma of testis converted to multiple myeloma].

A 55-year-old male was referred to our hospital with the chief complaint of painless right scrotal swelling. Serum tumor marker levels were all within the normal range. A right radical orchiectomy was performed and the histopathological diagnosis was plasmacytoma. Chest and abdominal computed tomographic scan and bone scintigraphy did not show metastasis. Serum and urine protein electrophoresis and bone marrow biopsy were negative. Diagnosis was primary testicular plasmacytoma (extramedullary plasmacytoma). Eleven months after the operation, multiple bone lesions appeared and our diagnosis was conversion to multiple myeloma.

Identification of genes responsible for reducing palladium ion in Escherichia coli.

Palladium (Pd) is commonly used as a catalyst for automobiles and electronic devices, and a reliable source of Pd is required for continued commercial applications. Biomineralization has attracted attention as an inexpensive and eco-friendly recycling approach for a continued supply of Pd. Escherichia coli is one of the best hosts for collecting Pd because it grows rapidly and requires an inexpensive minimal medium. Although E. coli can reduce Pd ions, the mechanism of reduction has not been thoroughly investigated. In this study, we investigated the genes involved in the reduction of Pd ions in E. coli. A gene responsible for the reduction of Pd ions was identified from approximately 4000 genes, other than essential genes, by using the single-gene-knockout library. The rate of reducing Pd ions by E. coli cells was evaluated. Among the investigated single-gene-knockout strains, 7 strains including the gene related to membrane transport, transcriptional regulation, and metabolic enzyme promote the reduction of Pd ions, and 73 strains including the genes related to formate metabolism and molybdopterin synthesis repress the reduction of Pd ions. Our results may provide a new perspective for the improvement of the bioreduction of minor metals.

A rare case of suspected intravascular large B-cell lymphoma forming nodular masses in the brain and around the prostate and seminal vesicle.

Intravascular large B-cell lymphoma (IVLBCL) is a rare subtype of diffuse large B-cell lymphoma (DLBCL). Furthermore, tumorigenesis is extremely rare. An 80-year-old man was admitted to our hospital with nervous symptoms. Imaging tests showed a brain tumor and mass lesions in the seminal vesicle and retroperitoneum. Transrectal biopsy of the seminal vesicle helped diagnose the patient with DLBCL. The patient's general status deteriorated rapidly, and he died on the 23rd day after admission. An autopsy was performed and the pathological diagnosis was DLBCL, specifically suspected as IVLBCL, with nodular masses in the brain and seminal vesicle.

Bioreductive deposition of platinum nanoparticles on the bacterium Shewanella algae.

An environmentally friendly method using the metal ion-reducing bacterium Shewanella algae was proposed to deposit platinum nanoparticles. Resting cells of S. algae were able to reduce aqueous PtCl(6)(2-) ions into elemental platinum at room temperature and neutral pH within 60min when lactate was provided as the electron donor. Biogenic platinum nanoparticles of about 5nm were located in the periplasm--a preferable, cell surface location for easy recovery of biogenic nanoparticles.

Direct determination of oxidation state of gold deposits in metal-reducing bacterium Shewanella algae using X-ray absorption near-edge structure spectroscopy (XANES).

X-ray absorption near-edge structure spectroscopy (XANES) was successfully employed to determine the gold valence in the metal-reducing bacterium Shewanella algae after exposure to a 1 mM aqueous HAuCl4 solution for 10-120 min. XANES spectra revealed the oxidation state of gold in the bacterial cells to be Au(0) without any contribution from Au(III), demonstrating that S. algae cells can reduce AuCl4- ions to elemental gold. Transmission electron microscopy (TEM) and energy dispersive X-ray (EDX) analysis confirmed that gold nanoparticles 5-15 nm in size were deposited in the periplasmic space of the bacterial cells; a preferable, cell surface location for the easy recovery of biogenic nanoparticles.

Cytotoxicity and colloidal behavior of polystyrene latex nanoparticles toward filamentous fungi in isotonic solutions.

The effects of surface physicochemical properties of functionalized polystyrene latex (PSL) nanoparticles (NPs) and model filamentous fungi Aspergillus oryzae and Aspergillus nidulans cultivated in different environment (aqueous and atmospheric environment) on the colloidal behavior and cytotoxicity were investigated in different isotonic solutions (154 mM NaCl and 292 mM sucrose). When the liquid cultivated fungal cells were exposed to positively charged PSL NPs in 154 mM NaCl solution, the NPs were taken into A. oryzae, but not A. nidulans. Atomic force microscopy revealed that the uptake of NPs was more readily through the cell wall of A. oryzae because of its relatively softer cell wall compared with A. nidulans. In contrast, the positively charged PSL NPs entirely covered the liquid cultivated fungal cell surfaces and induced cell death in 292 mM sucrose solution because of the stronger electrostatic attractive force between the cells and NPs compared with in 154 mM NaCl. When the agar cultivated fungal cells were exposed to the positively charged PSL NPs, both fungal cells did not take the NPs inside the cells. Contact angle measurement revealed that the hydrophobin on the agar cultivated cell surfaces inhibited the uptake of NPs because of its relatively more hydrophobic cell surface compared with the liquid cultivated cells.

Evaluation of the G145R Mutant of the Hepatitis B Virus as a Minor Strain in Mother-to-Child Transmission.

The role of the hepatitis B virus (HBV) mutant G145R, with a single change in amino acid 145 of the surface protein, as a minor population remains unknown in mother-to-child transmission. The minor strain as well as the major strain of the G145R mutant were evaluated in three cohorts using a locked nucleic acid probe-based real-time PCR. The breakthrough cohort consisted of children who were born to HBV carrier mothers and became HBV carriers despite immnoprophylaxis (n = 25). The control cohort consisted of HBV carriers who had no history of receiving the hepatitis B vaccine, hepatitis B immunoglobulin or antiviral treatment (n = 126). The pregnant cohort comprised pregnant women with chronic HBV infection (n = 31). In the breakthrough cohort, 6 showed positive PCR results (major, 2; minor, 4). In the control cohort, 13 showed positive PCR results (major, 0; minor, 13). HBeAg-positive patients were prone to have the G145R mutant as a minor population. Deep sequencing was performed in a total of 32 children (PCR positive, n = 13; negative, n = 19). In the breakthrough cohort, the frequency of the G145R mutant ranged from 0.54% to 6.58%. In the control cohort, the frequency of the G145R mutant ranged from 0.42% to 4.1%. Of the 31 pregnant women, 4 showed positive PCR results (major, n = 0; minor, n = 4). All of the pregnant women were positive for HBeAg and showed a high viral load. Three babies born to 3 pregnant women with the G145R mutant were evaluated. After the completion of immunoprophylaxis, 2 infants became negative for HBsAg. The remaining infant became negative for HBsAg after the first dose of HB vaccine. G145R was detected in one-fourth of the children with immunoprophylaxis failure. However, the pre-existence of the G145R mutant as a minor population in pregnant women does not always cause breakthrough infection in infants.

Estimation of the adhesive force distribution for the flagellar adhesion of Escherichia coli on a glass surface.

The effects of the presence or absence of microbial flagella and the microbial motility on the colloidal behaviors of microbial cells were quantitatively evaluated. The microbial cell attachment and detachment processes on a glass surface were observed directly using a parallel-plate flow chamber. Wild-type, flagellar paralyzed, and nonflagellated Escherichia coli strains were used as model microbial cells. In the cell attachment tests, the microbial adhesion rate in a 160mM NaCl solution was approximately 10 times higher than that in a 10mM solution, for all E. coli strains. The colloidal behavior of the microbial cells agreed well with the predictions of the DLVO theory. In addition, the microbial flagella and motility did not significantly affect the cell attachment, regardless of the existence of a potential barrier between the cell and the glass substratum. In the cell detachment tests, the cumulative number of microbial cells detached from the glass substratum with increasing flow rate was fit well with the Weibull distribution function. The list of strains arranged in order of increasing median drag force required to remove them was nonflagellated strain, flagellar paralyzed strain, and wild-type strain. These results indicated that the flagella and the flagellar motility inhibited the cell detachment from the glass substratum. Furthermore, a large external force would likely be required to inhibit the microbial adhesion in the early stage of the biofilm formation.

Intraoperative red cell salvage during obstetric surgery in 50 Japanese women.

OBJECTIVE: To determine the clinical usefulness of intraoperative cell salvage (ICS) in obstetrics. METHODS: A retrospective analysis was performed using data for 50 patients who had received ICS blood during obstetric surgery at 13 Japanese facilities between January 1, 2007 and December 31, 2013. The frequencies of ICS-associated adverse events, allogeneic blood transfusion (ABT), and preoperative autologous donation (PAD) were assessed. RESULTS: Placenta previa was the indication for ICS in 42 (84%) women. The ICS blood was reinfused in all women (median 366 mL; range 80 to at least 3715). No ICS-associated adverse events occurred. The median estimated blood loss (EBL) was 2171 mL (range 574-47 000); 27 (54%) women lost at least 2000 mL. ABT was not used in 33 (66%) women. Among 26 women who lost at least 2000 mL of blood and were included in analyses, 12 (44%) did not receive ABT. EBL was linearly correlated with the total volume of transfused blood (P<0.001). CONCLUSION: ICS caused no adverse events among women at elevated risk of peripartum hemorrhage and might be safe for use in obstetrics.

Measurement of microbial adhesive forces with a parallel plate flow chamber.

HYPOTHESIS: It was predicted that the colloidal behaviors of archaea and bacteria with disparate surface structure were different. In this study, the effects of the physicochemical properties of microbial cell surfaces on colloidal behavior were analyzed with Derjaguin-Landau-Verwey-Overbeek (DLVO) theory, thermodynamics, and powder technology. EXPERIMENTS: Cell attachment and detachment from model substrates were directly observed using a parallel plate flow chamber. Gram-negative Escherichia coli and archaeal Methanosarcina barkeri were used as model microbial cells, and positively and negatively charged glass slides were used as model substrates. FINDINGS: Microbial adhesion on both substrates agreed well with predictions calculated from DLVO theory, using experimental parameters. The total number of cells detached from the substrates as a function of flow rate was fit with the Weibull distribution function. In addition, the drag force required for detachment, which was estimated from the hydrodynamic forces, had a wide distribution; however, the forces became smaller with increasing ionic strength because of reduced electrostatic interactions between the cells and the substrate. M. barkeri could not be detached from positively charged substrates because it would entail a negative change in the interfacial energy of interaction. Thus adhesion was thermodynamically favored in this case.