Active site phosphoryl groups in the biphosphorylated phosphotransferase complex reveal dynamics in a millisecond time scale

AbstractThe N-terminal domain of Enzyme I (EIN) and phosphocarrier HPr can form a biphosphorylated complex when they are both phosphorylated by excess cellular phosphoenolpyruvate. Here we show that the electrostatic repulsion between the phosphoryl groups in the biphosphorylated complex results in characteristic dynamics at the active site in a millisecond time scale. The dynamics is localized to phospho-His15 and the stabilizing backbone amide groups of HPr, and does not impact on the phospho-His189 of EIN. The dynamics occurs with the kex of ∼500s−1 which compares to the phosphoryl transfer rate of ∼850s−1 between EIN and HPr. The conformational dynamics in HPr may be important for its phosphotransfer reactions with multiple partner proteins.Structured summary of protein interactionsEIN and HPr bind by nuclear magnetic resonance (View Interaction)

Proizvodnja antikomplementnih egzopolisaharida submerznim uzgojem gljive Flammulina velutipes

Seven species of basidiomycetes have been investigated for anti-complementary activity in hot water extracts and ethanol soluble fractions. Since Flammulina velutipes had the most potent activity, culture conditions for its mycelial growth were optimized to increase the production efficiency of anti-complementary exopolysaccharides. The optimal medium composition was (in g/L): galactose 15, sodium nitrate 5, glutamic acid 3, KH2PO4 2.5 and MgSO4·7H2O 0.6. Optimal production of anti-complementary activity was achieved at pH=3.5–5.5 and 25 °C. With these optimal medium and culture conditions, mycelial dry mass was maximized at 3.17 mg/mL after 6 days of cultivation in a 5-liter stirred-tank bioreactor, without pH control. The anti-complementary activity of exopolysaccharides increased sharply after 4 days of cultivation, and showed a high level at 5–6 days of cultivation. A periodate-labile carbohydrate moiety played a leading role in the anti-complementary activity exhibited by exopolysaccharide produced from F. velutipes. Results of tests on the anti-complementary activity in the absence of Ca²+ and immunoelectrophoresis indicated that the mode of complement activation by exopolysaccharide from F. velutipes is via both the classical and alternative pathways and that the activation degree is almost the same in each pathway.Istražena je antikomplementna aktivnost spojeva ekstrahiranih vrućom vodom i etanolom iz sedam vrsta gljiva stapčara. Optimirani su uvjeti uzgoja micelija gljive s najvećom aktivnosti, Flammulina velutipes, radi povećanja proizvodnje antikomplementnih egzopolisaharida. Optimalni sastav podloge bio je (u g/L): galaktoza 15, natrijev nitrat 5, glutamična kiselina 3, KH2PO4 2,5 i MgSO4·7H2O 0,6. Optimalna proizvodnja postignuta je pri pH=3,5-5,5 i 25 ºC. Pri tim uvjetima proizvedena je maksimalna količina suhe tvari od 3,17 mg/L nakon 6 dana uzgoja u bioreaktoru s miješalicom zapremnine 5 L, bez kontrole pH-vrijednosti. Antikomplementna aktivnost egzopolisaharida naglo se povećala nakon 4 dana, te je bila visoka nakon 5-6 dana uzgoja. Šećerni je ostatak, podložan djelovanju perjodata, glavni razlog antikomplementne aktivnosti egzopolisaharida gljive F. velutipes. Rezultati testiranja takve aktivnosti u odsutnosti Ca²+ iona i imunoelektroforeza upućuju na to da se komplementi egzopolisaharida iz F. velutipes aktiviraju klasičnim i alternativnim putem te da je stupanj aktivacije skoro jednak za oba puta

Proizvodnja antikomplementnih egzopolisaharida submerznim uzgojem gljive Flammulina velutipes

Seven species of basidiomycetes have been investigated for anti-complementary activity in hot water extracts and ethanol soluble fractions. Since Flammulina velutipes had the most potent activity, culture conditions for its mycelial growth were optimized to increase the production efficiency of anti-complementary exopolysaccharides. The optimal medium composition was (in g/L): galactose 15, sodium nitrate 5, glutamic acid 3, KH2PO4 2.5 and MgSO4·7H2O 0.6. Optimal production of anti-complementary activity was achieved at pH=3.5–5.5 and 25 °C. With these optimal medium and culture conditions, mycelial dry mass was maximized at 3.17 mg/mL after 6 days of cultivation in a 5-liter stirred-tank bioreactor, without pH control. The anti-complementary activity of exopolysaccharides increased sharply after 4 days of cultivation, and showed a high level at 5–6 days of cultivation. A periodate-labile carbohydrate moiety played a leading role in the anti-complementary activity exhibited by exopolysaccharide produced from F. velutipes. Results of tests on the anti-complementary activity in the absence of Ca²+ and immunoelectrophoresis indicated that the mode of complement activation by exopolysaccharide from F. velutipes is via both the classical and alternative pathways and that the activation degree is almost the same in each pathway.Istražena je antikomplementna aktivnost spojeva ekstrahiranih vrućom vodom i etanolom iz sedam vrsta gljiva stapčara. Optimirani su uvjeti uzgoja micelija gljive s najvećom aktivnosti, Flammulina velutipes, radi povećanja proizvodnje antikomplementnih egzopolisaharida. Optimalni sastav podloge bio je (u g/L): galaktoza 15, natrijev nitrat 5, glutamična kiselina 3, KH2PO4 2,5 i MgSO4·7H2O 0,6. Optimalna proizvodnja postignuta je pri pH=3,5-5,5 i 25 ºC. Pri tim uvjetima proizvedena je maksimalna količina suhe tvari od 3,17 mg/L nakon 6 dana uzgoja u bioreaktoru s miješalicom zapremnine 5 L, bez kontrole pH-vrijednosti. Antikomplementna aktivnost egzopolisaharida naglo se povećala nakon 4 dana, te je bila visoka nakon 5-6 dana uzgoja. Šećerni je ostatak, podložan djelovanju perjodata, glavni razlog antikomplementne aktivnosti egzopolisaharida gljive F. velutipes. Rezultati testiranja takve aktivnosti u odsutnosti Ca²+ iona i imunoelektroforeza upućuju na to da se komplementi egzopolisaharida iz F. velutipes aktiviraju klasičnim i alternativnim putem te da je stupanj aktivacije skoro jednak za oba puta

Incidence and risk factors of subsyndromal delirium after curative resection of gastric cancer

Background: Subsyndromal delirium, a condition in which patients exhibit some, but not all, of the symptoms of delirium, can negatively affect the outcomes of patients with cancer. However, the incidence of subsyndromal delirium in patients with gastric cancer is unknown. Here, we investigated the incidence and risk factors of subsyndromal delirium after curative resection of gastric cancer. Methods: We recruited consecutive patients with gastric cancer who were scheduled for curative resection at a tertiary hospital. Patients' subsyndromal delirium symptoms were serially assessed preoperatively and 1, 2, 3, and 7 days postoperatively using the Delirium Rating Scale-Revised-98 (DRS-R-98). A DRS-R-98 score of 8-14 at any postoperative assessment was considered to indicate subsyndromal delirium. Sociodemographic and pre-/intraoperative clinical data were also assessed. Logistic regression analyses were used to determine the associated risk factors. Results: Data were analysed from 163 out of 217 eligible patients. Postoperative delirium occurred in one patient (0.6%) and subsyndromal delirium occurred in 19 patients (11.7%). Age >= 70 years (odds ratio, [OR] 3.85; 95% confidence interval [0], 136-10.92; p = 0.011) and education level <= 9 years (OR, 3.98; 95% CI, 139-11.41; p= 0.010) were independent risk factors of subsyndromal delirium after adjusting for preoperative cognitive function. Other pre-/intra-operative variables including anxiety/depression, poor sleep quality, and anaesthesia duration were not associated with subsyndromal delirium. Conclusions: In contrast to the low incidence of delirium among patients undergoing curative resection of gastric cancer, a substantial proportion of such patients experienced subsyndromal delirium. Considering the prognostic implications, more careful detection and management of subsyndromal delirium may be warranted in patients with gastric cance

Higher Lesion Detection by 3.0T MRI in Patient with Transient Global Amnesia

PURPOSE: Transient global amnesia (TGA) patients were retrospectively reviewed to determine the usefulness of high-field strength MRI in detecting probable ischemic lesions in TGA. MATERIALS AND METHODS: We investigated the lesion detection rate in patients with TGA using 1.5T and 3.0T MRI. Acute probable ischemic lesions were defined as regions of high-signal intensity in diffusion weighted image with corresponding low-signal intensity in apparent diffusion coefficient map. RESULTS: 3.0T MRI showed 11 out of 32 patients with probable ischemic lesions in the hippocampus with mean lesion size of 2.8 +/- 0.6 mm, whereas 1.5T MRI detected no lesion in any of 11 patients. There were no significant differences in clinical characteristics between the groups of 1.5 and 3.0T MRI. CONCLUSION: High-field strength MRI has a higher detection rate of probable ischemic lesions than low-field strength MRI in patients with TGA.ope

Characteristics of Outpatients with Pandemic H1N1/09 Influenza in a Tertiary Care University Hospital in Korea

The pandemic H1N1/09 emerged rapidly in Korea. Here, we describe the clinical characteristics of outpatients in Seoul, Korea who were infected in the 2009 H1N1 pandemic. We reviewed the cases of outpatients with pandemic H1N1/09 who visited a tertiary care teaching hospital between September 1 and December 31, 2009. Infection with pandemic H1N1/09 was confirmed by molecular tests. Of a total of 7,182 tests, 3,020 (42.0%) were positive. Compared with 473 cases of influenza-like illness (ILI), the 586 confirmed cases of pandemic H1N1/09 differed in age [odds ratio (OR) 0.975] and fulfilling at least one of the following factors: age <5 or ≥65 years, history of contact with other pandemic H1N1/09-infected individuals (OR 0.611), fever ≥37.8℃ (OR 3.567), cough (OR 2.290), and myalgia (OR 1.559). The sensitivity of the best criteria, "fever (≥37.8℃) plus cough" (41.03%) in this study was lower than that of the Korea Centers for Disease Control and Prevention (KCDC) criteria (47.95%), whereas the positive likelihood ratio (3.55) and positive predictive value (81.6) of this criteria was higher than those of the KCDC criteria (2.98 and 78.7, respectively). The clinical characteristics of pandemic H1N1/09 are, in many regards, indistinguishable from those of ILI. Moreover, the accuracy and predictability of criteria which include only symptoms or signs were not sufficient to diagnose pandemic H1N1/09 infection. Therefore, use of a combination of symptoms with confirmatory laboratory testing is necessary for accurate diagnosis of pandemic H1N1/09