Interleukin-like EMT Inducer (ILEI, FAM3C) in Melanoma Partial Phenotype Switching

Interleukin like EMT-Inducer (ILEI, FAM3C) is a secreted factor that contributes to the epithelial-to-mesenchymal transition (EMT), a cell biological process that confers metastatic properties to a tumor cell. Initially, we found that ILEI mRNA is highly expressed in melanoma metastases but not in primary tumors, suggesting that ILEI contributes to the malignant properties of melanoma. While melanoma is not an epithelial cell-derived tumor and does not undergo a traditional EMT, melanoma undergoes a similar process known as phenotype switching in which high MITF (micropthalmia-related transcription factor) expressing proliferative cells (MITF-high) switch to a low expressing invasive state (MITF-low). We observed that MITF-high proliferative cells express low levels of ILEI (ILEI-low) and MITF-low invasive cells express high levels of ILEI (ILEI-high). Next, we used in vitro and in vivo assays to show that knockdown of ILEI attenuates invasive potential but does not affect MITF expression or chemoresistance. We used gene expression analysis to show that ILEI regulates several genes involved in the MITF-low invasive phenotype including JARID1B (KDM5B), HIF-2 (EPAS1), and BDNF. Gene set enrichment analysis suggested that ILEI-regulated genes are enriched for JUN signaling, a known regulator of the MITF-low invasive phenotype. Additionally, we found that inducing phenotype switching towards the MITF-low invasive state increases ILEI mRNA expression, whereas phenotype switching towards the MITF-high proliferative state decreases ILEI mRNA expression. Mechanistically, we found that the transcription factor upstream stimulatory factor 1 (USF1) regulates FAM3C transcription. In conclusion, we demonstrate that phenotype switching regulates ILEI expression through USF1, and that ILEI in turn regulates the invasive potential of MITF-low melanoma cells

Use of Procalcitonin in Patients on Chronic Hemodialysis: Procalcitonin Is Not Related with Increased Serum Calcitonin

Objectives. To investigate whether procalcitonin (PCT) could be useful for detecting bacterial infections in patients on hemodialysis (HD) and with increased calcitonin (CT). Methods. This prospective study included 42 males and 34 females on HD. The infection group consisted of 15 patients with proven bacterial infections; the other 61 patients were designated as the noninfection group. Serum C-reactive protein (CRP), interleukin (IL)-6, white blood cell (WBC) count, immature and total neutrophil (I/T) ratio, and CT were measured at the beginning of HD, and serum PCT levels at the beginning of HD and after HD. Results. The mean CT level in the both groups was apparently higher than that of nonchronic kidney disease. Significantly higher values between the infection and noninfection groups were seen for CRP, IL-6, WBC, I/T ratio, PCT, and CT. The PCT value of the area under the receiver operating characteristic curve was 0.921, which was significantly higher than the values for CRP (0.853; P < 0.01), IL-6 (0.739; P < 0.01), WBC (0.692; P < 0.01), and I/T ratio (0.584; P < 0.01). Conclusions. PCT was useful marker of bacterial infection in patients on HD and with increased CT. PCT levels should be determined before HD

Magnetic resonance (MR) imaging assessment for glossopharyngeal neuralgia : value of three-dimensional T2-reversed MR imaging (3D-T2R) in conjunction with other modes of 3D MR imaging

Background: To retrospectively examine the usefulness of gray-scale reversal imaging of T2-weighted images (3D-T2R) in conjunction with other modes of 3D MRI for preoperative assessments in patients with glossopharyngeal neuralgia (GPN) due to neurovascular compression. Material/Methods: Imaging findings on 3D-T2R, constructive interference in steady state (CISS), and MRA were analyzed with reference to operative charts in 10 patients with GPN. Results: Offending vessels were associated with the posterior inferior cerebellar artery (PICA) in 9 of 10 patients (90%). Eight of the 10 patients (80%) had offending vessels located at the supraolivary fossette. Of those eight patients, six (75%) had a shift of the ipsilateral vertebral artery to the affected side. Five (42%) and seven (48%) contact points were associated with the root entry/exit zone and the peripheral nerve system segment, respectively. In six of nine contact points (67%), 3D-T2R demonstrated the pathomorphological features at the contact points better than CISS. Conclusions: The offending vessels were mostly associated with posterior inferior cerebellar arteries, were frequently located at the supraolivary fossette, and had attachments at the root entry/exit zone and at the peripheral segment of the glossopharyngeal nerve, which was well demonstrated on 3D-T2R

Hydrogenase-3 Contributes to Anaerobic Acid Resistance of Escherichia coli

BACKGROUND: Hydrogen production by fermenting bacteria such as Escherichia coli offers a potential source of hydrogen biofuel. Because H(2) production involves consumption of 2H(+), hydrogenase expression is likely to involve pH response and regulation. Hydrogenase consumption of protons in E. coli has been implicated in acid resistance, the ability to survive exposure to acid levels (pH 2-2.5) that are three pH units lower than the pH limit of growth (pH 5-6). Enhanced survival in acid enables a larger infective inoculum to pass through the stomach and colonize the intestine. Most acid resistance mechanisms have been defined using aerobic cultures, but the use of anaerobic cultures will reveal novel acid resistance mechanisms. METHODS AND PRINCIPAL FINDINGS: We analyzed the pH regulation of bacterial hydrogenases in live cultures of E. coli K-12 W3110. During anaerobic growth in the range of pH 5 to 6.5, E. coli expresses three hydrogenase isoenzymes that reversibly oxidize H(2) to 2H(+). Anoxic conditions were used to determine which of the hydrogenase complexes contribute to acid resistance, measured as the survival of cultures grown at pH 5.5 without aeration and exposed for 2 hours at pH 2 or at pH 2.5. Survival of all strains in extreme acid was significantly lower in low oxygen than for aerated cultures. Deletion of hyc (Hyd-3) decreased anoxic acid survival 3-fold at pH 2.5, and 20-fold at pH 2, but had no effect on acid survival with aeration. Deletion of hyb (Hyd-2) did not significantly affect acid survival. The pH-dependence of H(2) production and consumption was tested using a H(2)-specific Clark-type electrode. Hyd-3-dependent H(2) production was increased 70-fold from pH 6.5 to 5.5, whereas Hyd-2-dependent H(2) consumption was maximal at alkaline pH. H(2) production, was unaffected by a shift in external or internal pH. H(2) production was associated with hycE expression levels as a function of external pH. CONCLUSIONS: Anaerobic growing cultures of E. coli generate H(2) via Hyd-3 at low external pH, and consume H(2) via Hyd-2 at high external pH. Hyd-3 proton conversion to H(2) is required for acid resistance in anaerobic cultures of E. coli

NPK Fertilizers for Elephant Foot Yam (Amorphophallus Paeoniifolius (Dennst.) Nicolson) Intercropped with Coffee Trees

Fertilizer application in elephant foot yams (Amorphophallus paeoniifolius (Dennst.) Nicolson) intercropping system is rare in Indonesia, therefore, NPK fertilizers experiment was conducted under the shade of 10-year-old coffee plantation at Leuwikopo Experimental Farm, Bogor, Indonesia, in order to increase the productivity of elephant foot yam intercropped with coffee trees. Prior to planting, 20 ton ha-1 of goat manure was applied. Four NPK combinations, i.e., N, P2O5, K2O at the rate of 0, 0 and 0; 100, 60 and 80; 125, 60 and 100; and 150, 60 and 120 kg ha-1, were applied. Results showed that there were no significant differences in leaf number per plant, petiole size and rachis length among treatments. Application of NPK decreased photosynthetic rates, while increasing rate of N and K2O had no effect on photosynthetic rates. NPK application at the 100 N, 60 P2O5 and 80 K2O kg ha-1 (N100P60K80 treatment) or larger prolonged growth duration regardless of NPK levels, and there was a close relationship between corm yield and growth duration. As a result, corm fresh mass was higher in the 100:60:80 kg ha-1 treatment than in control. In the N125P60K100 and N150P60K120 kg ha-1 treatments, leaves were damaged by heavy rains and winds, counteracting beneficial effect of NPK on growth duration and corm yield. These results suggested the importance of delay of entering dormancy for an increase in productivity of A. paeoniifolius

Storage and allogeneic transplantation of peripheral nerve using a green tea polyphenol solution in a canine model

<p>Abstract</p> <p>Background</p> <p>In our previous study, allogeneic-transplanted peripheral nerve segments preserved for one month in a polyphenol solution at 4°C could regenerate nerves in rodents demonstrated the same extent of nerve regeneration as isogeneic fresh nerve grafts. The present study investigated whether the same results could be obtained in a canine model.</p> <p>Methods</p> <p>A sciatic nerve was harvested from a male beagle dog, divided into fascicules of < 1.5 mm diameter, and stored in a polyphenol solution (1 mg/ml) for one month at 4°C. The nerve fascicles were transplanted into 10 female beagle dogs to bridge 3-cm right ulnar nerve gaps. In the left ulnar nerve in each dog, a 3-cm nerve segment was harvested, turned in the opposite direction, and sutured in situ. Starting one day before transplantation, the immunosuppressant FK506 was administered subcutaneously at doses of 0.1 mg/kg daily in four dogs (PA0.1 group), 0.05 mg/kg daily in four dogs (PA0.05 group), or 0.05 mg/kg every other day in two dogs (PA0.025 group). Twelve weeks after surgery, electrophysiological and morphological studies were performed to assess the regeneration of the right and left ulnar nerves. The data for the right ulnar nerve were expressed as percentages relative to the left ulnar nerve. Polymerase chain reaction (PCR) was used to identify the sex-determining region of the Y-chromosome (<it>Sry</it>) and β-actin to investigate whether cells of donor origin remained in the allogeneic nerve segments. FK506 concentration was measured in blood samples taken before the animals were killed.</p> <p>Results</p> <p>The total myelinated axon numbers and amplitudes of the muscle action potentials correlated significantly with the blood FK506 concentration. Few axons were observed in the allogeneic-transplanted nerve segments in the PA0.025 group. PCR showed clear <it>Sry</it>-specific bands in specimens from the PA0.1 and PA0.05 groups but not from the PA0.025 group.</p> <p>Conclusions</p> <p>Successful nerve regeneration was observed in the polyphenol-treated nerve allografts when transplanted in association with a therapeutic dose of FK506. The data indicate that polyphenols can protect nerve tissue from ischemic damage for one month; however, the effects of immune suppression seem insufficient to permit allogeneic transplantation of peripheral nerves in a canine model.</p